Previous studies recorded the prevalence of gastrointestinal nematodes (GIN) in Limpopo Province. However, the studies did not address the seasonal patterns of infection and did not cover all districts of Limpopo Province, namely; Capricorn, Sekhukhune, Waterberg, Mopani, and Vhembe. It is, therefore, important to provide up to date information on the prevalence and seasonal occurrence data of GIN in all districts of Limpopo province. The present study was conducted to determine the occurrence of anthelmintic resistance (AR) and document the prevalence of GIN infecting sheep in five districts of Limpopo Province, South Africa. Forty animals in each district were used for fecal egg count reduction test (FECRT) to determine AR against ivermectin (0.2 mg/kg), levamisole (LEV) (5 mg/kg), and albendazole (7.5 mg/kg). Egg hatch test (EHT) was used to determine AR against thiabendazole (TBZ) and micro-agar larval development test (MALDT) was used for both TBZ and LEV. Naturally, infected sheep (n=780) were sampled for prevalence across five districts of Limpopo. FAMACHA© eye-color score estimations were also performed for each study animal. FECRT showed occurrence of AR in most of the districts and a few with suspected resistance. EHT results showed AR development against TBZ for all districts, while the MALDT showed no AR against LEV in all districts, but detected AR against TBZ in Sekhukhune, Capricorn, and Waterberg. Haemonchus contortus was the most resistant species. A high nematode prevalence (88-100%) and 1210-1861 eggs per gram (EPG) was observed in all districts during the hot wet season, decreasing to 75-80% (453-1202 EPG) during the cold dry season. The sheep revealed a FAMACHA© mean score of 3, indicating mild anemia during the hot wet season except for Vhembe district that revealed a FAMACHA© mean score of 4 during the hot wet season, indicating anemia. AR recorded in Limpopo Province may be due to under-dosing caused by lack of weighing equipment and high treatment frequencies due to lack of proper training on anthelmintic use. The detection of AR in Limpopo is an important finding because it will help in outlining effective management systems against GIN. Keywords: anthelmintic resistance, fecal egg counts, FAMACHA, seasonal prevalence, sheep.

Zinc has a wide spectrum of biological activities and its deficiency has been related to various dysfunctions. This study aimed to clarify the clinical, hematological, and biochemical changes in Holstein dairy calves with naturally occurring hypozincemia before and after treatment. This study was carried out on 25 Holstein dairy calves <1 month of age in the El-Salhya Dairy Farm, Al-Sharqiya Province, Egypt. Calves were born from apparent healthy dams without any clinical signs of zinc deficiency. They were divided into two groups. The first group (G1) included five clinically healthy calves that were used as controls. The second group (G2) included 20 calves suffering from alopecia and skin lesions. The diseased calves were then treated by oral administration of zinc oxide at the rate of 80 mg/day for 10 successive days and then 20 mg/week for 2 weeks (G3). A total of 90 samples, whole blood and serum samples were collected during the study across all groups. Whole blood was evaluated for complete blood count and serum was used to estimate total protein, albumin, globulin, zinc, calcium, magnesium, phosphorus, and the activity of alkaline phosphatase (ALP) and aspartate aminotransferase. The diseased calves had macrocytic normochromic anemia. Total leukocytes, neutrophils, and lymphocytes were significantly reduced in the diseased calves than in the control and treated groups. Biochemical analysis of serum revealed a highly significant decrease in the globulin, zinc, and calcium concentrations in the diseased calves than in the control and treated groups. ALP activity was significantly lower in the diseased and treated calves than in control. There were no differences in any other parameters between the groups. Zinc deficiency naturally occurring in calves caused clinical, hematological, and biochemical alterations such as alopecia, skin abnormalities, and macrocytic normochromic anemia. In addition, zinc deficiency altered the cell-mediated immunity as indicated by leukopenia and lymphopenia. These alterations were improved by oral administration of zinc oxide. Keywords: alopecia, calf, complete blood count, skin lesions, zinc deficiency.

Although the production of safe food for human consumption is the primary purpose for animal rearing, the environment and well-being of the animals must also be taken into consideration. Based on microbiological point of view, the production of healthy food from animals involves considering foodborne pathogens, on the one hand and on the other hand, the methods used to fight against germs during breeding. The conventional method to control or prevent bacterial infections in farming is the use antibiotics. However, the banning of these compounds as growth promoters caused many changes in animal breeding and their use has since been limited to the treatment and prevention of bacterial infections. In this function, their importance no longer needs to be demonstrated, but unfortunately, their excessive and abusive use have led to a double problem which can have harmful consequences on consumer health: Resistance to antibiotics and the presence of antibiotic residues in food. The use of probiotics appears to be a suitable alternative to overcome these problems because of their ability to modulate the immune system and intestinal microflora, and further considering their antagonistic role against certain pathogenic bacteria and their ability to play the role of growth factor (sometimes associated with prebiotics) when used as feed additives. This review aims to highlight some of the negative effects of the use of antibiotics in animal rearing as well as emphasize the current knowledge on the use of probiotics as a feed additive, their influence on animal production and their potential utility as an alternative to conventional antibiotics, particularly in poultry, pig, and fish farming.

Postpartum uterine infectious diseases, such as pyometra, have discrepancy with both health and, subsequently, productivity of dairy cows due to its high prevalence and the high cost of treatment. Therefore, this study investigates the influence of pyometra on the reproductive indices, the metabolic profile, and oxidant/antioxidant parameters of the pyometric animal compared to those of healthy ones. The study included 30 cows. The animals were differentiated into two groups of 15 cows each: A group of pyometra and a control group. All pyometric cows were subjected to breeding soundness examination after the end of pyometra and were compared to the control group. Blood samples were obtained to assess the levels of glucose, non-esterified fatty acids (NEFA), triglycerides (TGs), cholesterol, albumin, total protein, alanine aminotransferase, aspartate aminotransferase (AST), alkaline phosphatase (ALP), blood urea nitrogen (BUN), creatinine, calcium (Ca), phosphorus, sodium, potassium, progesterone hormone (P4), malondialdehyde (MDA), total antioxidant capacity (TAC), glutathione peroxidase (GPx), and superoxide dismutase. Results revealed significant prolonged duration of first estrus, the days open, and the required number of services due to pyometra. The pyometra group yielded increased levels of NEFA, TGs, ALP, BUN, creatinine, MDA, and progesterone hormone. In addition, significant decrease in the levels of glucose, cholesterol, albumin, Ca, phosphorus, sodium, TAC, GPx, and superoxide dismutase was observed in the pyometra group. Finally, no difference in the concentrations of total protein, ALT, AST, and potassium was observed in the pyometra group. The reproductive indices was adversely influenced in cows with postpartum pyometra, and metabolic profile, involving energy balance signals and liver function indicators, revealed differences between the two groups. Increased levels of oxidative stress parameters and decrease levels of antioxidant levels were also found, suggesting that pyometra is an incentive for oxidative stress. Overall, checking the energy balance, metabolic imbalances, and oxidant/antioxidant profile, accompanied with pre-emptive procedures during the postpartum period, is essential and can reduce the chances of such diseases and possible noxious results in highly productive cows. Keywords: dairy cows, metabolic profile, oxidant/antioxidant parameters, postpartum pyometra, reproductive indices.

Goats are reared for their meat, mohair and other socio-cultural needs in Lesotho. Helminth infections are some of the major setbacks in the goat production industry due to their negative impact on animals' health, resulting in significant losses on meat and mohair production and death. A cross-sectional study was conducted to determine the prevalence, fecal egg infestation, and morphological identification of gastrointestinal parasites in goats. Fecal samples were collected from 765 goats and subjected to McMaster egg counting techniques using the flotation method. Statistical analyses were performed using the Statistical Package for the Social Sciences (SPSS v.26.0). The overall prevalence of gastrointestinal parasites was 94.7%, and the identified gastrointestinal parasites were nematodes (64.7%), coccidia (25.8%), and cestodes (4.2%). Haemonchus contortus was identified as the prevalent gastrointestinal nematode species found in goats. The prevalence and fecal egg count of gastrointestinal parasites were significantly higher (p<0.05) in goats located in the highlands and Senqu River Valley, while goats in the lowlands demonstrated a significantly (p<0.05) higher prevalence of H. contortus. Immature goats and kids were more significantly (p<0.05) prone to gastrointestinal parasites. The nematodes and coccidia infestations were prevalent in goats located in the highlands and foothills, respectively, whereas nematode and coccidia fecal egg loads were higher in goats located in the foothills and Senqu River Valley, respectively. Keywords: fecal egg counts, gastrointestinal parasites, goats, Haemonchus contortus, prevalence.

Toxocara vitulorum is a bovine intestinal nematode. Immune pictures following infection are conflicting and stopping anthelmintic albendazole treatment recording reversed liver abnormalities. The purpose of this work was to evaluate the therapeutic potential of bone marrow mesenchymal stem cells (BMMSCs) therapy, subsequent to albendazole administration in rats infected with T. vitulorum. The ultrasonographic and histopathological examinations as well as serum liver enzymes activity and the kinetics of recovery were investigated. The correlation of cell-mediated and humoral immune pictures was assessed by assaying immunoglobulins, splenocytes viability, phagocytic index, and Th1/Th2 cytokines. The cultured BMMSCs counting were 4.21×104 cells/cm2 with 96.03% viability. Flow-cytometric analysis indicated positive CD90 (82%), CD105 (79%) and negative CD34 (0.37%), CD45 (0.42%), attesting to the suitability of the isolated BMMSCs for use in therapy. Transplantation of BMMSCs after albendazole administration significantly reduced the release of liver enzymes (p<0.05) indicating liver cellularity improvement. The ultrasonographic, macroscopic, and histopathological findings confirmed the biochemical results. Significant elevation in the levels of tumor necrosis factor (TNF)-α and interferon (INF)-γ with a decline in interleukin (IL)-4 was observed in the untreated model (p<0.05). However, albendazole treatment followed by BMMSCs therapy significantly lowered the release of TNF-α and INF-γ, associated with significant production of IL-4 and IL-10 (p<0.05). The final results indicated that the liver functions, histopathological findings, and immune parameters were aggravated after experimental T. vitulorum infection. Albendazole treatment followed by BMMSCs therapy was found to assist in regeneration of injured hepatic tissue. Besides, it appeared to modulate host defensive immune responses against T. vitulorum antigens. This work could define more clearly the events that manipulate the host immune, histopathological, and biochemical responses to minimize obstacles in using stem cell therapy in animal toxocariosis. Keywords: albendazole, bone marrow mesenchymal stem cells, cell therapy, rat, Toxocara vitulorum.

This review article delineates the role of chelated minerals and phytogenic feed additives (PFAs) cocktail supplementation in improving the overall health status and production performance of poultry birds and its economic effects in the poultry industry. Organically complexed minerals have many advantages over inorganic sources. It has improved absorption and efficacy, which meets the bird's requirements comfortably with a low-dose level. Hence, inorganic forms can be replaced with lower-dose levels of organic minerals without any adverse effects on production performances in broilers and layers. PFAs possess medicinal properties, such as antimicrobial, antioxidant, adaptogenic, and immunomodulatory, therefore, could be recommended as supplements. They are also growth promoters that enhance the overall health status and augment poultry birds' production performance. Furthermore, the tremendous potential of PFAs could be extracted with the recent advances in science and technology. With the advantages of organically complexed minerals and multiple beneficial applications, there is a resurgence to develop PFAs as a cocktail of organic minerals to improve the overall health status of poultry birds and augment their productivity, which, in turn, helps the poultry industry to grow decisively and economically.

Backyard pigs farming is a rearing system associated with poor hygienic and sanitary conditions of the pig, often causing public health and food safety problems. Therefore, this study was conducted to investigate the occurrence of potentially zoonotic parasites in population pig reared under backyard farming in the Bucaramanga metropolitan area, Northeast Colombia. From September to December 2019, a total of 558 fecal samples from 64 backyard pig farms were examined for the presence of enteric protozoan infection. The coprological diagnosis was done by direct examination using Lugol's iodine solution, buffered saline solution, and Kinyoun technique. In addition, blood samples were collected from 200 pigs. Serum was collected and used for the detection of Trichinella spiralis and Taenia solium cysticercosis infections, using the enzyme-linked immunosorbent assay. The overall prevalence of zoonotic protozoa in the Bucaramanga metropolitan area was 65.2%, reporting 52.7% prevalence for Balantidium coli, 33.7% for Entamoeba coli, and 5.7% for Cryptosporidium spp. Regarding the prevalence by municipalities, there was no statistical association (p>0.05), indicating that the prevalence was similar in the region under study. Pigs >7 months of age showed to be a risk factor for B. coli infection, indicating that the prevalence increases with the age, while pigs raised at >1000 masl and access to latrines, increased infection risk for E. coli and Cryptosporidium spp. infections. In the present study, T. spiralis infection was not detected in the analyzed sera, while T. solium cysticercosis infection was found to be 40.5%. The high prevalence of protozoan infections and porcine cysticercosis reported in this study could be due to poor facilities, and lack of hygiene in the facilities, and suggests the possible transmission of these parasite populations between pigs and humans, thus increasing the transmission of parasites zoonotic potential. Therefore, appropriate sanitary management practices and deworming programs should be adopted to reduce the prevalence of these infectious agents. Keywords: Colombia, cysticercosis, parasite, pigs, Trichinella, zoonotic.

Ultrasonography (USG) is useful for non-invasively identifying changes that occur in soft tissue architecture. The objective of this research was to demonstrate postpartum (PP) uterine involution through the changes of perineal muscle intensity and thickness in Madura beef cow by ultrasonography. Madura's breed cows used in the research consist of eight non-pregnant (NP) cows and three PP cow. The transrectal and transperineal USG imaging of NP cows was performed on days 1, 33, and 65. USG imaging of PP cows was performed every day starting from day 1 (24 h after parturition) until day 21 PP. Transrectal USG of the reproductive tract was performed for the cervix, corpus uteri, and cornua uteri. USG was performed transcutaneously over the perineal area for coccygeus and levator ani muscles at the longitudinal and transverse angles. Reproductive tract diameter and perineal muscle intensity and thickness were measured with ultrasound imaging. The analysis of the sonogram of PP cows showed that the diameter of the cervix, corpus uteri, and cornua uteri decreased within 21 days PP. The transverse view of the coccygeus muscle of PP cows showed decreased muscle intensity and thickness. On the other hand, the longitudinal view showed increased coccygeus muscle intensity and thickness. The transverse view of the coccygeus muscle of NP cows showed increased muscle intensity, while muscle thickness was reduced. Sonogram analysis of the levator ani muscle of PP cows showed decreased muscle intensity with increasing muscle thickness. However, imaging of the levator ani muscle of NP cows showed a decrease in both intensity and muscle thickness. There was a significant difference in the mean value intensity of the scanning view analysis results of the levator ani muscle of the PP cow (523.6 AU increased to 672.1 AU) and the NP cow (515.9 AU decreased to 465.4 AU). Furthermore, there was a significant difference (p<0.05) in the mean value thickness of both scanning view analyses of the coccygeus and levator ani muscles of PP cows (5.8 mm increased to 6.5 mm and 3.8 mm increased to 4.8 mm, respectively) and NP cows (8.8 mm increased to 9.1 mm and 5.9 mm decreased to 4.9 mm, respectively). We found that the perineal muscles, namely, the levator ani muscle and coccygeus muscle, change in size, and intensity during uterine involution as demonstrated on Madura beef cow. Keywords: coccygeus, levator ani, Madura beef cow, postpartum, ultrasonography.

The anesthetic-surgical stress response consists of metabolic, neuroendocrine, hemodynamic, immunological, and behavioral adaptations through chemical mediators such as the adrenocorticotropic hormone, growth hormone, antidiuretic hormone, cortisol, aldosterone, angiotensin II, thyroid-stimulating hormone, thyroxine, triiodothyronine, follicle-stimulating hormone, luteinizing hormone, catecholamines, insulin, interleukin (IL)-1, IL-6, tumor necrosis factor-alpha, and prostaglandin E-2. Behavioral changes include adopting the so-called prayer posture, altered facial expressions, hyporexia or anorexia, drowsiness, sleep disorders, restriction of movement, licking or biting the injured area, and vocalizations. Overall, these changes are essential mechanisms to counteract harmful stimuli. However, if uncontrolled surgical stress persists, recovery time may be prolonged, along with increased susceptibility to infections in the post-operative period. This review discusses the neurobiology and most relevant organic responses to pain and anesthetic-surgical stress in dogs and cats. It highlights the role of stress biomarkers and their influence on autonomous and demeanor aspects and emphasizes the importance of understanding and correlating all factors to provide a more accurate assessment of pain and animal welfare in dogs and cats throughout the surgical process.

Feline leukemia virus (FeLV) is classified as Retroviridae gammaretrovirus. FeLV occurs worldwide, including Malaysia. Thus far, only one decade-old study on molecular characterization of Malaysian FeLV isolates exists, which resulted in a scarcity of updated information of current FeLV isolates circulating in Malaysia. This study was conducted to determine the status of FeLV in clinically ill cats and to study the molecular characterization and phylogenetic relatedness of the current isolates. Convenience sampling was performed in 20 cats from the Gasing Veterinary Hospital in Selangor. Plasma and saliva samples were collected from 15 clinically ill cats and 5 healthy cats subjected to one-step reverse transcription-polymerase chain reaction with primers targeting a highly conserved gene of U3-LTR-gag. Two clinically ill cats' plasma and saliva samples tested positive for FeLV RNA. Partial nucleotide sequencing and phylogenetic analysis revealed that the current isolates were 94-99% homologous to the previous Malaysian and Japanese FeLV isolates. Current FeLV isolates from this study displayed higher similarity with the previous Malaysian isolates, signifying that a similar FeLV strain circulated among the cat population in Selangor. Keywords: feline leukemia virus, Malaysia, one-step reverse transcription polymerase chain reaction, phylogenetic analysis.

Blood collection from dogs is the most commonly performed procedure in the medical clinic. However, different factors can interfere with the quality of the material collected, potentially causing complications for patients. Simulated skill training is a teaching strategy designed to provide early training to students, develop their skills and self-confidence, and increase the procedure's success while reducing complications. Therefore, the aim of this study was to evaluate skill training using a low-fidelity simulator in the peripheral venipuncture procedure and examine the training's influence on the in vivo procedure. To assess skill training, this study used a low-fidelity simulator in the peripheral venipuncture procedure and examines the training's effect on the in vivo procedure. In total, 100 dogs, 65 undergraduate students, 3 veterinarians, and 4 previously trained evaluators participated. The canine in vivo venipuncture procedure was evaluated both before and after the simulated skill training and the low-fidelity simulator training. Data were collected on participants' self-confidence levels. Local complications occurred during in vivo practice; however, after training, they decreased. Gloves were more frequently used during the procedure, resulting in a reduction of both harvest attempts and complications, as well as increased levels of self-confidence in post-training participants. The simulator developed had low fidelity, low cost, and was easy to create. Skill training in peripheral venipuncture using a low-fidelity simulator positively influences student learning, increases their self-confidence during in vivo harvesting, and reduces the complications of the procedure, improving patient well-being. Keywords: canine, clinical skills training, evaluation, self-confidence, veterinary simulation.

Giardia duodenalis is one of the most common enteric protozoan parasites in vertebrates, such as humans, domestic and wild animals, causing giardiasis. To the best of our knowledge, little is known about the genetic diversity of G. duodenalis assemblages. This study aimed to identify genetic diversity of G. duodenalis assemblages in Iranian stray dogs. A total of 450 fecal samples were collected from 2015 to 2016 from stray dogs of Northwest Iran. All specimens were observed microscopically following concentration and flotation techniques. Subsequently, DNA samples were extracted, amplified, and sequenced targeting the glutamate dehydrogenase gene. The overall prevalence of G. duodenalis in infected dogs was estimated at 1.6%, based on microscopic and molecular diagnoses. Sequencing and phylogenetic analyses indicated a high level of genetic diversity of assemblage C (haplotype diversity; 0.802). The pairwise sequence distances between the identified isolates of assemblage C showed an intradiversity of 0.3%-1.3% and identity of 98.7%-100%. Current findings indicate that a significant genetic diversity of G. duodenalis assemblage C haplotypes is unequivocally circulates among stray dogs in Northwest Iran. Keywords: genetic variation, Giardia lamblia assemblage C, glutamate dehydrogenase, Iran, stray dogs.

Poultry is becoming an increasingly important source of protein in the Nepalese diet. The Chitwan region of Nepal is the hub of the emerging poultry industry. Little is known about the prevalence of non-typhoidal Salmonella (NTS) on poultry farms or the role of farm management practices that may contribute to the presence of NTS on farms. The role of poultry in the transmission of Salmonella enterica to humans is also poorly defined. This descriptive study seeks establish baseline data through estimation of the prevalence of NTS on broiler and layer operations in various farms of the Chitwan district of Nepal. Based on district documents on poultry production and meat marketing, a purposive sampling of 18 commercial poultry farms comprising ten broilers farms and eight layers farms was conducted. Environmental samples including water, litter, feces, feed, farm, and eggshell swabs were randomly collected from each farm. Samples were cultured and tested for the presence of NTS; positives were serotyped, and antimicrobial susceptibility determined. A comprehensive farm and practice questionnaire was administered to each farm manager. The farm level point prevalence rate was 55% (10 of 18 farms) for S. enterica. Of the total 288 farm environmental samples collected, 26 samples (9%) were positive. The rate of isolation varied according to the origin of samples: Water (27.5%), feces (10.6%), litter (8.6%), farm swabs (5%), feed (1.8%), and eggshells (0%). Farm management variables/risk factors are summarized and categorized as non-modifiable and modifiable for analysis. Broiler operations were more likely to be positive than layer operations as were poultry houses with two or less open sides. All-in/all-out management style was found to be protective. Due to the small sample size (18 farms), no associations reached statistical significance. Based on environmental sampling results, NTS is highly prevalent on the poultry farms in the Chitwan district of Nepal. Certain risk factors are associated with finding NTS on farms. Our findings are generally in agreement with other studies in similar countries with rapidly emerging poultry industries. The identification of risk factors provides owners, technicians, and veterinarians with some guidance to help reduce the prevalence of NTS on farms. This baseline data are critical to understanding the epidemiology of zoonotic strain of NTS in the region and are necessary for the design of future studies and mitigation plans and underlines the need for a one-health approach to protect public health-related to Salmonella spp. from poultry farms. Keywords: farm risk factors, Nepal, non-typhoidal Salmonella, poultry, Salmonella enterica.

Antibiotic-resistant infections are one of the leading threats to public health globally. Indiscriminate use of antibiotics in food animal production is an important driver of resistance, particularly among foodborne pathogens such as non-typhoidal Salmonella enterica (NTS). While there has been extensive research on antimicrobial-resistant (AMR) S. enterica in India and China, there have been few studies in countries in South Asia, including Nepal. This is particularly important with the rise of commercial poultry farming in Nepal as a means of economic development and nutritional subsistence. This descriptive study seeks to identify the prevalence and resistance patterns of NTS serotypes focusing on Chitwan, Nepal's leading poultry producing district. A mixture of purposive and judgment sampling of 18 poultry farms and 20 slaughterhouses representing a broad geographic distribution across multiple municipalities in Chitwan was conducted in May 2019. Environmental samples taken from poultry farms included: Water, litter, feces, feed, farm swabs, and eggshell swabs. Biological samples taken from nearby slaughterhouses included: Muscle, heart, liver, skin, cecum, crop, and spleen. Samples were cultured and tested for the presence of NTS. Positive isolates were serotyped and tested for antimicrobial susceptibility to seven antibiotics known to be important to both human and animal health regionally. Farm practices were also characterized through a survey, the results of which are detailed in the accompanying paper. Out of 708 samples (288 environmental and 420 biological), 103 (15%) tested positive for NTS (9% of environmental; n=26, 18% of biological; n=77). The percentage of positive environmental and biological samples varied by source. Environmental sample positive rates were water (27.5%), feces (10.6%), litter (8.6%), farm swabs (5%), feed (1.8%), and eggshells (0%). Biological sample positive rates were skin (28%), heart (23%), crop (20%), muscle (15%), liver (15%), spleen (15%), and cecum (12%). Out of 103 positive S. enterica isolates, 48.5% were identified as Salmonella Typhimurium, 35% Salmonella Enteritidis, 7.8% Salmonella Gallinarum, 4.9% Salmonella Virchow, and 3.9% were Salmonella Agona. Of the 103 positive isolates, 80 (78%) were resistant to at least one antibiotic, and 21 (20%) were multidrug-resistant (MDR). NTS is highly prevalent among Chitwan's growing poultry industry with higher rates of positivity found in slaughterhouse samples compared with environmental samples from farms. In addition, a high rate of AMR (78%) was revealed, and an extremely concerning number of those were shown to be MDR (20%). This baseline data has important implications for poultry production and consumption in the region. Further research will elucidate the extent to which this contamination and drug resistance is impacting the health of the local population and help inform treatment and management strategies. Note: The characterization of the poultry industry and practices that might be linked to NTS contamination in the Chitwan district are detailed in the previous paper in this series (www.veterinaryworld.org/Vol.14/February-2021/14.pdf). Keywords: antibiotic resistance, Nepal, non-typhoidal Salmonella, poultry, serotype.

Several free-roaming dogs can be easily found in the public areas of Bali. They go out in search of foods and friends and defecate everywhere. In general, these groups of dogs do not receive good healthcare from their owners and are generally threatened by some disease-causing organisms, especially helminths. This study was conducted to identify and measure the prevalence of soil-transmitted helminths (STHs) that cause infection in free-roaming dogs in Bali Province, Indonesia. A total of 1611 fresh dog fecal samples were collected from all areas of Bali Province and subjected to qualitative fecal examination using flotation techniques to obtain STH eggs. The incidence of STHs in free-roaming dogs was 38.36%. However, three types of STHs were identified, including Ancylostoma spp., Ascaris spp., and Trichuris spp. Ancylostoma spp. had the highest prevalence of 37.8%, followed by Ascaris spp. and Trichuris spp. at 6.02% and 0.87%, respectively. Multiple infections of these worms were also recorded. The polyparasitism prevalence of Ancylostoma spp. and Ascaris spp. was 3.85%, followed by that of Ancylostoma spp. and Trichuris spp. at 0.5% and that of Ascaris spp. and Trichuris spp. at 0.06%. There were no altitude-wise differences in the prevalence of STH infection. The prevalence of STHs was high in free-roaming dogs. This finding necessitates more serious attention as it affects both animal and public health. Keywords: Ancylostoma spp., Ascaris spp., Bali, distribution, free-roam dogs, prevalence, soil-transmitted helminth, Trichuris spp.

Several factors had been concerned with the developmental competence of the sheep oocyte. This study aims to investigate the effect of adding growth factors (insulin-like growth factor 1 [IGF-1] and epidermal growth factor [EGF]) in the maturation medium of ewe oocytes selected based on brilliant cresyl blue (BCB) screening on in vitro maturation (IVM), fertilization, and pre-implantation embryo development. Cumulus-oocyte complexes (COCs) were obtained from the ovaries of slaughtered ewes by either aspiration or slicing techniques. COCs were in vitro matured in a medium containing IGF-1 and EGF (control group). For BCB screening, oocytes were stained and divided into BCB+ oocytes that matured in the same maturation conditions without adding growth factors (Group 2) or in the presence of growth factors (Group 3), and BCB– oocytes that matured in medium without growth factors (Group 4) or with growth factors (Group 5). The supplementation of the maturation medium with growth factors during IVM of (BCB+) oocytes resulted in a significant increase in nuclear maturation rate (90.9%), fertilization rate (75.6%), and embryo developmental rates (60.0%, 46.7%, and 33.3% for cleavage, morula, and blastocyst, respectively). Culturing BCB+ oocytes in a maturation medium containing both EGF and IGF-1 showed a significant improvement in nuclear maturation, fertilization, and pre-implantation embryo development in vitro. Keywords: brilliant cresyl blue, embryo development, growth factors, In vitro fertilization, sheep.

Anacyclus pyrethrum (L.) has been used in traditional North African and Indian medicine for the treatment of several diseases such as cancer, rheumatism, epilepsy, diabetes, and Alzheimer's disease. Despite its medical benefits, few studies have examined its toxicity. The present study evaluated the acute toxicity of hydroethanolic extracts of different parts (roots, seeds, leaves, and capitula) of two varieties of A. pyrethrum (L.), namely, A. pyrethrum var. pyrethrum (L) and A. pyrethrum var. depressus (Ball) Maire, in mice. Acute toxicity was evaluated after the oral administration of different extracts at doses of 300, 500, and 2000 mg/kg. Mortality, body weight, general behavior, and adverse effects were observed daily for 14 days. At the end of the experiment, mice were sacrificed, and biochemical parameters and histopathology of the liver, kidneys, and spleen were analyzed. The extracts of different parts of both plants induced no signs of toxicity or mortality during the observation period, excluding capitulum and seed extracts, which induced slight sedation at a dose of 2000 mg/kg. The LD50 of the extracts was estimated to exceed 2000 mg/kg. The administration of A. pyrethrum var. pyrethrum roots at a dose of 300 mg/kg resulted in significantly increased AST levels. However, the A. pyrethrum var. depressus root extract induced significant increases in the levels of both transaminases (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]). The remaining extracts of both plants at a dose of 500 mg/kg significantly increased AST levels. Moreover, all plant extracts excluding the A. pyrethrum var. pyrethrum capitulum extract at 2000 mg/kg provoked significant increases in AST levels, and A. pyrethrum var. depressus roots provoked a significant increase of ALT levels. Meanwhile, mice treated with high doses of extracts (2000 mg/kg) displayed histopathological changes in the liver, kidneys, and spleen characterized by hepatic distress, inflammatory infiltration, focal tubular necrosis, vascular congestion, and lymphoid hyperplasia. The results of the present study indicate that the hydroethanolic extracts of different parts of two varieties of A. pyrethrum (L.) were not toxic in mice at low concentrations, whereas some toxic effects were detected in mice treated at 2000 mg/kg. Keywords: acute oral toxicity, Anacyclus pyrethrum var. depressus (Ball) Maire, Anacyclus pyrethrum var. pyrethrum (L), histopathology, serum biochemistry analysis.

Sarcocystosis is a zoonotic infection caused by various species of Sarcocystis organisms with a worldwide geographic distribution. This study investigated the presence of Sarcocystis organisms in cattle and sheep slaughtered at an abattoir in Sulaimani Province in North Iraq. A total of 130 muscle samples were collected during May, June, and July of 2020, including 80 samples from sheep and 50 samples from cattle. Samples were examined visually for macrosarcocysts. The peptic digestion method was used to analyze fresh muscle tissue samples for detecting microsarcocysts followed by microscopic examination. Furthermore, muscle samples were fixed and stained with hematoxylin and eosin for histopathological examination. In the gross examination, macroscopic cysts were not detected in both cattle and sheep; hence, all the prevalence data were obtained through microscopic observation of muscle samples. The peptic digestion method revealed the presence of banana-shaped bradyzoites in 90% and 92.5% of slaughtered cattle and sheep muscle samples, respectively. Organ-wise prevalence revealed that 95% and 92% of esophageal samples of sheep and cattle contained Sarcocystis spp., respectively. Moreover, 90% and 88% of sheep and cattle diaphragms were respectively infected. Histopathological examination of tissue sections revealed two morphologically distinct types of microsarcocysts, including thin-walled and thick-walled, in both sheep and cattle. The suspected Sarcocystis spp. were Sarcocystis tenella and Sarcocystis arieticanis in sheep and Sarcocystis cruzi and Sarcocystis bovifelis or Sarcocystis hominis in cattle. Infective stages of different Sarcocystis spp. are widespread in the study area environment. Keywords: cattle, muscle samples, prevalence rate, Sarcocystis, sheep.

Sheep productivity in developing countries is crucial, as this animal is an essential source of meat and wool. Myostatin (MSTN) plays an important role in the regulation of muscle mass through the regulation of muscle growth, differentiation, and regeneration. The present study sought to investigate genetic variation in the first intron of the MSTN gene and the association of variants with growth traits in major sheep breeds in Egypt (Barki, Ossimi, and Rahmani) and Saudi Arabia (Najdi) using polymerase chain reaction (PCR) and sequencing. Blood samples were collected, and DNA was extracted from 75 animals. A 386 bp fragment in the first intron of the MSTN gene was amplified using PCR. Polymorphic sites were detected using direct sequencing and then correlated with growth traits using a general linear model. Sequence analysis of the first intron of MSTN gene identified six single-nucleotide polymorphisms (SNPs) in the studied breeds. Four mutual SNPs were determined: c.18 G>T, c.241 T>C, c.243 G>A, and c.259 G>T. In addition, two SNPs c.159 A>T and c.173 T>G were monomorphic (AA and TT, respectively) in the Ossimi, Rahmani, and Najdi breeds and polymorphic in the Barki breed. The association analysis revealed that the c.18 G>T and c.241 C>T significantly associated (p<0.05) with birth weight and average daily weight gain, respectively. Our results strongly support MSTN as a candidate gene for marker-assisted selection in sheep breeding programs. Furthermore, the identified variants may be considered as putative markers to improve growth traits in sheep. Keywords: growth traits, Myostatin gene, polymorphism, sheep.

Clostridium perfringens is one of the multiple drug-resistant intestinal pathogens causing necrotic enteritis disease, leading to great economic losses in poultry farms. This study aimed to evaluate the potential use of peppermint oil and its microemulsion (ME) as an alternative to antibiotics to control necrotic enteritis in broiler chickens. Peppermint oil ME formulation (15% oil/water) was prepared and characterized by zeta potential, Fourier transform infrared, high-resolution transmission electron microscopy, and liquid chromatography–mass spectrometry (LC-MS/MS). The minimal inhibitory concentrations of the peppermint oil and its ME were investigated. A total of 80 commercial one day old Arbor Acres broiler chickens were randomly assigned to four groups of 20 birds each. The four groups were the negative control, positive control, peppermint oil (0.5 mL/mL water/10 days old), and its ME (0.25 mL/mL water/10 days old) groups. C. perfringens was orally provided at concentration of 1×108 CFU/mL on days 14, 15, and 16. Clinical signs and mortality were observed daily. Growth performance, gross lesions and cecal samples were investigated and examined on days 21, 28, and 35. Peppermint oil ME formulation has a polydispersity index, zeta potential and droplet size of 0.234, –24 mV±4.19, and 29.96±1.56 nm, respectively. LC–MS/MS analysis of oil and ME revealed common presence of phenolic compounds such as rosmorinic (360.31 g/mol), chlorogenic acid (354.31 g/mol), hesperidin (610.56 g/mol), and luteolin 7-O-β- glucuronide (462.1 g/mol). The treated groups with peppermint oil and ME showed lower lesions, mortality and colony-forming units in addition to higher growth performance (p<0.05) compared to the positive control group. Our study suggests the potential efficacy of peppermint oil and ME in the reduction of necrotic enteritis lesions and C. perfringens count. Keywords: broiler chicken, feed conversion ratio, microemulsion, nanotechnology, necrotic enteritis, peppermint oil.

Lumbosacral intervertebral disk disease (IVDD) in cats usually develops concurrent with constipation, spondylosis deformans, and sacralization. However, the prevalence of lumbar IVDD in cats was considered low, and there was less information on the incidence of non-traumatic lumbosacral vertebral abnormalities that may affect large bowel dysfunction. This study aimed to retrospectively investigate the relationship between non-traumatic lumbosacral vertebral abnormalities, both congenital and acquired, and large bowel dysfunction in cats. Of 3108 cats that were presented to the Diagnostic Imaging Unit, the Small Animal Teaching Hospital, Faculty of Veterinary Science, Chulalongkorn University, between March 2016 and February 2018, 1365 cats met the inclusion criteria. All abdominal radiographs were reviewed, and all subsequent data were recorded, including the presence of congenital or acquired lumbosacral lesions, number of lumbar vertebrae, and length of the second, fifth, and last lumbar vertebrae, including the type of lumbar abnormalities. Moreover, radiographic information relating to constipation and megacolon was also collected. Non-traumatic lumbosacral vertebral abnormalities were observed in 29.74% of cats. The most common congenital lumbosacral vertebral abnormalities were six lumbar vertebrae, sacralization, and lumbarization, whereas most common acquired lumbosacral abnormalities were bone spur, narrowing disk space, spondylosis deformans, and lumbosacral degeneration, respectively. Cats with abnormal lumbosacral vertebrae are prone to have more problems with the large bowel (p=0.0057; odds ratio=1.731). Moreover, congenital and acquired lumbosacral abnormalities were also at risk of large bowel abnormalities (p=0.0069; odds ratio=1.920 and p<0.0001; odds ratio=4.107, respectively). This study revealed the evidence and distribution of the variation in feline lumbar anatomy and also elucidated that cats with abnormal lumbar vertebral columns were more likely to have problems with distal gastrointestinal tracts than those without. Keywords: cat, constipation, large bowel, lumbosacral, vertebrae.

Garcinia kola seed (GKS) is used to prevent and cure a number of gastric-related ailments. GKS contain a copious amount of polyphenols and can be utilized as a natural growth promoter in the nutrition of weaned rabbits. This study aimed to determine the dietary effects of GKS meal (GKSM) on the growth performance, hematology and serum biochemical parameters of weaned rabbits. GKS were dried and ground into powder. Thirty-two 8-week-old crossbred rabbits with an average weight of 614 g were randomly divided into four dietary groups. The diets were designated as follows: Control (corn-soybean based diet with 0% GKSM) and the GKSM-supplemented diets with 1.5% GKSM, 3% GKSM, and 4.5% GKSM. On the 56th day of the feeding trial, blood was collected from the marginal ear vein of all rabbits and transferred into two separate labeled tubes. The first set of blood was used to determine the hematological indices. The second set of blood was transferred into plain bottles and allowed to coagulate. The coagulated blood was subjected to standard methods of serum separation, and the sera were harvested and used to evaluate serum biochemical parameters. Although the average final body weight was highest in rabbits fed with 1.5% GKSM; this value was similar to rabbits fed with 0% and 4.5% GKSM. The average daily weight gain was highest in rabbits fed with 1.5% GKSM, while the feed conversion ratio was improved in the 0% and 1.5% GKSM groups. The dietary treatments also had a significant effect on the red blood cell count (RBC) and hemoglobin (Hb) concentration, while other blood parameters did not differ significantly (p>0.05). Higher inclusion levels (3% and 4.5%) of GKSM led to a significant increase in RBC and Hb values (p<0.05). The total protein increased at all levels of GKSM inclusion (p<0.05). Bilirubin, sodium, and potassium levels significantly decreased at 4.5% GKSM inclusion (p<0.05). Urea levels were lowered at 1.5% and 4.5% GKSM inclusion, while cholesterol levels were decreased at 3% and 4.5% dietary levels. From the results of the present study, the supplementation of up to 4.5% GKSM revealed no harmful effect on the hematological and serum biochemical parameters of weaned rabbits, while their growth performance improved at a 1.5% inclusion level of GKSM. Keywords: Garcinia kola, performance, production, rabbits, supplementation.

Adipose tissue-derived stromal vascular fraction (SVF) contains a heterogeneous cell population comprising multipotent adipose-derived stem cells. Regenerative therapy using adipose-derived SVF has broad applications in bone tissue engineering due to the superior osteogenic potential of SVF. This study was designed to standardize and characterize adipose-derived SVF obtained from New Zealand white rabbits for bone tissue engineering and other potential applications. Ten skeletally mature and clinically healthy adult New Zealand white rabbits were used in this study. The SVF was prepared using surgically resected interscapular adipose tissue following enzymatic digestion with 0.1% collagenase type I solution. The SVF pellet obtained after the final centrifugation step was suspended in a 0.5 mL control solution to obtain ready-to-use adipose-derived SVF. The freshly prepared SVF was characterized based on the total SVF cell count and cell yield per gram of adipose tissue. The SVF cells were enumerated using a hemocytometer. Interscapular adipose tissue depots are ideal for preparing autologous adipose-derived SVF due to the ease of access. The interscapular adipose-derived SVF prepared by enzymatic digestion had an average cell yield of 3.15±0.09×106 cells/g adipose tissue. Freshly prepared SVF had a total cell count of 3.15±0.09×104 cells/μL. The enzymatic digestion of adipose tissue using 0.1% collagenase resulted in better cell yield per gram than methods previously reported in rabbits. The use of adipose-derived SVF can preclude the requirement for an additional culture period. In addition, it may also reduce the risk of extensive cell contamination, which makes it a safe and cost-effective strategy for bone tissue engineering applications. Keywords: adipose tissue, characterization, New Zealand white rabbit, standardization, stromal vascular fraction.

Marker-assisted selection has many advantages over conventional selection in animal breeding. The candidate gene approach has been applied to identify genetic markers associated with economically important traits in livestock. This study was established to investigate variation in the hormone-sensitive lipase (HSL) and leptin receptor (LEPR) genes, and their association with growth traits in Barki lambs. Records for birth weight (BW), pre-weaning average daily gain (ADG1), weaning weight (WW), post-weaning average daily gain (ADG2), and marketing weight (MW) were obtained from 247 Barki lambs. Polymerase chain reaction–single-stranded conformational polymorphism analyses were used to detect variation in exon 9 of HSL and exon 19 of LEPR. General linear models were used to test for associations between the variation in ovine HSL and LEPR, and growth traits. The SSCP banding patterns for HSL showed three variants (H1, H2, and H3), which contained two nucleotide-sequence differences (c.1865C>T and c.2038T>C). Two SSCP banding patterns (L1 and L2) were observed for LEPR and these contained two nucleotide-sequence differences (c.2800G>A and c.2978C>G). The HSL genotype showed no effect on the studied traits. The LEPR genotype was proven to have significant effects (p<0.05) on ADG2 and MW. The presence of the L1 variant was associated (p<0.01) with decreased ADG2 and MW. The finding of an association between LEPR gene variation and growth rate after weaning in Barki lambs warrants efforts to improve this trait. Keywords: Barki lambs, growth traits, hormone-sensitive lipase, leptin receptor.

Probiotics improve intestinal balance through bacterial antagonism and competitive exclusion. This study aimed to investigate the in vitro antimicrobial activity, as well as the in vivo preventive, immunological, productive, and histopathological modifications produced by probiotic Bacillus subtilis. The in vitro antimicrobial activities of B. subtilis (5×106 CFU/g; 0.5, 1.0*, 1.5, and 2.0 g/L) were tested against Escherichia coli O157: H7, Salmonella Typhimurium, Candida albicans, and Trichophyton mentagrophytes after exposure times of 0.25, 0.5, 1, and 2 h using minimal inhibitory concentration procedures. A total of 320 1-day-old female Ross broiler chickens were divided into five groups. Four out of the five groups were supplemented with 0.5, 1.0*, 1.5, and 2.0 g/L probiotic B. subtilis from the age of 1 day old. Supplemented 14-day-old broiler chickens were challenged with only E. coli O157: H7 (4.5×1012 CFU/mL) and S. Typhimurium (1.2×107 CFU/mL). A total of 2461 samples (256 microbial-probiotic mixtures, 315 sera, 315 duodenal swabs, and 1575 organs) were collected. The in vitro results revealed highly significant (p<0.001) killing rates at all-time points in 2.0 g/L B. subtilis: 99.9%, 90.0%, 95.6%, and 98.8% against E. coli, S. Typhimurium, C. albicans, and T. mentagrophytes, respectively. Broilers supplemented with 1.5 and 2.0 g/L B. subtilis revealed highly significant increases (p<0.01) in body weights, weight gains, carcass weights, edible organs' weights, immune organs' weights, biochemical profile, and immunoglobulin concentrations, as well as highly significant declines (p<0.01) in total bacterial, Enterobacteriaceae, and Salmonella counts. Histopathological photomicrographs revealed pronounced improvements and near-normal pictures of the livers and hearts of broilers with lymphoid hyperplasia in the bursa of Fabricius, thymus, and spleen after supplementation with 2.0 g/L B. subtilis. The studies revealed that 1.5-2.0 g of probiotic B. subtilis at a concentration of 5×106 CFU/g/L water was able to improve performance, enhance immunity, and tissue architecture, and produce direct antimicrobial actions. Keywords: broiler chickens, histopathological photomicrographs, immunity, in vitro antimicrobial, in vivo preventive, probiotics.

Cordyceps militaris (CM) is a fungus that has been used to enhance aphrodisiac activity in men, but to date, no studies have focused on its antidiabetic properties. This study aimed to investigate the effects of CM on reproductive performance of streptozotocin (STZ)-induced diabetic male rats. Six-week-old Wistar rats were randomly divided into four groups: control Group 1 consisting of healthy rats; Group 2, healthy rats treated with CM (100 mg/kg); Group 3, diabetic untreated rats; and Group 4, diabetic rats treated with CM (100 mg/kg). Rats were orally administered with vehicle or CM for 21 days. The body weight, blood glucose level, food intake, epididymal sperm parameter, sexual behavior, serum testosterone level, and antioxidant parameters were determined. The results indicated that CM treatment in STZ-induced diabetic rats significantly improved the epididymal sperm parameter and serum testosterone level and, in turn, their copulatory behavior. CM treatment in diabetic rats significantly ameliorated malondialdehyde level and significantly improved the glutathione and catalase levels. These results provide new information on the pharmacological properties of CM in ameliorating testicular damage due to oxidative stress and improving sexual performance in diabetic male rats. Keywords: Cordyceps militaris, diabetes mellitus, sexual behavior, sperm, streptozotocin.

Elephant endotheliotropic herpesvirus (EEHV) is a serious disease, threatening the life of young elephants. Many elephants have been infected with no clinical signs and may serve as carriers spreading this disease. It is important to monitor the disease through clinical signs and molecular diagnosis. In this study we investigated the occurrence of EEHV and the efficiency of different techniques used to monitor EEHV infection in various samples and populations of Asian elephants. Blood and trunk swabs were collected from live elephants, while visceral organs (lung, digestive tract, spleen, lymph nodes, and kidney) were collected from dead elephants. EEHV was detected by polymerase chain reaction (PCR) in whole blood, trunk swabs, and visceral organs as samples, while elephant anti-EEHV immunoglobulin G (IgG) in serum was detected by enzyme-linked immunosorbent assay (ELISA). A total of 162 samples were analyzed in this study: 129 from healthy, 26 from dead, and 7 from sick elephants. The present study showed that the overall incidence of EEHV was 40.1% (n=65/162). Approximately 46.2% (n=12/26) and 85.7% (n=6/7) of dead and sick elephants were positive for EEHV by PCR, respectively. All sick elephants that were young and affected by EEHV clinical disease tested negative for the IgG antibody ELISA, suggesting primary EEHV infection in this group. In addition, 2.3% (n=3/129) of subclinical infections were detected using PCR, and trunk swab samples showed slightly higher sensitivity (5.3%, n=2/38) to detect EEHV than whole blood (1.2%, n=1/84). As many as, 48.4% (n=44/91) of healthy elephants were EEHV seropositive (ELISA-positive), suggesting that many elephants in Thailand had previously been infected. Overall, 30% of dead wild elephants had been infected with EEHV (n=3/10). Moreover, statistical analysis revealed no significant differences in the EEHV detection rate between different age groups or sexes (p>0.05). PCR is better than ELISA to detect EEHV active infection in dead/sick elephants and to monitor EEHV in young elephants. ELISA is suitable for detecting previous EEHV infection and carriers, particularly adults. Theoretically, we could use both PCR and ELISA to increase the sensitivity of testing, along with observing abnormal behavior to efficiently monitor this disease. Identification of EEHV carriers within elephant populations is important to prevent transmission to healthy individuals, especially young elephants with high mortality from EEHV. This is the first report from Thailand regarding EEHV infection in wild elephants, showing the importance of preventing disease transmission between captive and wild elephants. Keywords: Carrier, elephant endotheliotropic herpesvirus, elephant, enzyme-linked immunosorbent assay, polymerase chain reaction.