Clenbuterol as a β2-agonist drug was investigated according to the concentration of the drug available in the bodies of goats and according to the level of sensitivity of the instruments used for detection. The objective of the current study was to determine withdrawal times after giving a therapeutic dose that resulted in safe slaughters. Five healthy male goats with a mean body weight of 20.64 kg were treated with a single dose of 5.10-3 mg/kg in the BW onto jugular vein. Whole blood samples of approximately 5 mL were taken in a time series at 5, 30, 60, 90, 150, 210, 270, 390, 510, 630, and 750 min. At 24 h posttreatment, all subjects were sacrificed, and 300 g samples of the liver were obtained. The plasma concentration and liver residue of the drug were observed by reverse-phase high-performance liquid chromatography. The drug reached a maximum concentration of 19.233±0.331 μg/mL at 5 min, and the elimination half-life was at 173.25 min. The limit detection was obtained at 0.053 μg/mL. A one-way analysis of variance between all goats showed that elimination of the clenbuterol in their bodies was similar (p=1.00), with a withdrawal time of 1,479.326 min and no residues in the liver (p<0.05). Safe times for slaughter were determined to be at 2 days, 13 h, and 12 min as the 2nd safety factor (SF) time and 3 days, 1 h, and 58 min as the 3rd SF time with the liver organ free from residue. Keywords: elimination half-life, new method for calculating withdrawal time, prescriptions for obtained β2-agonist, residues in liver.

The aim of this study was to evaluate the relationship of scrotal circumference (SC) with plasma testosterone, seminal vesicles (SVs) weight, and its secretion as measurable indicators of fertility and also to sequence and establish phylogenetic relatedness of certain SV protein genes with other species as such integrated approach is lacking. Altogether, 59 apparently healthy male buffaloes sacrificed at slaughterhouse were selected (irrespective of breed) for measuring SC and collecting blood and paired SVs. The SC was measured at greater curvature using soft thread. In the present study, blood plasma testosterone, cholesterol, protein, and glucose in addition to SV fructose, citric acid and proteins in SV fluid were also estimated. The SV tissue was fixed in RNAlater for RNA extraction.Male buffaloes were categorized as per total SV weight into Group I (<5.0 g), Group II (5.0-7.84 g), and Group III (>8.0 g) and dentitions-I (≤18 months), II (18-24 months), and III (≥24 months) to assess the effect of weight and dentition age on SC, SV weight, and its certain secretions. Data were analyzed using linear model procedure including Tukey HSD test and Pearson's correlation coefficient. Variance inflation and condition index were also used to assess multicollinearity. Gross and histomorphological evaluation of SVs did not show any abnormality. Macronutrients (plasma protein, glucose, and cholesterol) showed non-significant (p>0.05) variation between groups. The SC and SV weight varied significantly (p<0.05) with a significant positive relationship with plasma testosterone, SV protein, fructose, and citric acid. In addition, testosterone concentration also showed increasing trend from Groups I to III but increased significantly (p<0.05) from Group II to III with positive and significant correlations with SV protein, fructose, and citric acid similar to SV weight and SC. Binders of sperm protein (BSP1, 3, and 5) genes (full length) were sequenced and established an evolutionary relationship which is lacking in buffalo. The present findings established a significant positive correlation of SC with that of other fertility parameters related to SVs weight and its secretions: Fructose, citric acid, and protein (inclusive of BSPs sequenced full length), and testosterone. Therefore, the present integrated approach along with certain semen quality attributes reflecting epididymis function could be used as a predictive fertility marker for grading and selection of breeding bulls and their progenies to develop outstanding bull mother farm. Keywords: male buffalo, morphology, scrotal circumference, seminal vesicles, sequencing, testosterone.

Brucellosis is a major constraint to livestock production in Egypt as well as many developing countries worldwide. Bovine brucellosis is an economically important disease with reproductive failure as a principal manifestation resulting in abortion, premature birth and decreased milk production in females, and orchitis and epididymitis in males. In spite of the efforts of Egyptian veterinary services to overcome brucellosis, the disease is still prevalent in both animals and humans and represents one of the most important public health hazards in Egypt. The aim of the present work was to investigate the efficacy of the control program implemented by the General Organization of Veterinary Services in Brucella infected buffalo farm on serological, molecular, cultural, and histopathological basis. Brucella melitensis biovar 3 was recovered from 6 buffalo-cows. Blood samples were collected from a total of 750 non-vaccinated lactating buffalo-cows. These animals were proved positive for Brucella by the Egyptian brucellosis national program. Sera were tested using buffered acidified plate antigen test and rose Bengal test as screening tests and complement fixation test as a confirmatory test. Positive animals were separated for slaughtering under the supervision of the Egyptian veterinary authorities. Remaining animals were tested every 3 weeks with slaughtering of positive cases and this continued until the remaining animals revealed three successive negative serological tests. Different lymph nodes (prescapular, prefemoral, mediastinal, retropharyngeal, and supramammary) were collected from 11 Brucella seropositive buffalo-cows slaughtered after being confirmed serologically as Brucella infected cases. Samples were collected and processed for bacterial isolation and nucleic acid detection using polymerase chain reaction (PCR). Parts of these specimens were fixed in 10% neutral buffered formalin for 48 h then processed by paraffin embedding technique. "Test and slaughter" policy was applied on Brucella infected dairy buffalo farm. The program continued for 6 months with slaughtering of positive cases until the herd was proved Brucella free. B. melitensis biovar 3 could be recovered from six buffalo-cows. Universal PCR confirmed Brucella on genus level and Bruce-ladder multiplex, PCR confirmed the presence of B. melitensis on the species level. Histopathological examination of Brucella-infected lymph nodes revealed massive rarified and depleted lymphoid areas of both sub-capsular and deep cortical lymphoid follicles, macrophage cells granulomatous reaction, as well as fat, infiltrates, and chronic vasculitis. The chronic nature of Brucella lesions has been confirmed in this study as indicated by the chronic vasculitis and collagen deposition. Freedom status from brucellosis in this study required 6 months which are considered long time allowing the spread of infection to other localities especially under unhygienic conditions, husbandry system favoring mixed populations of different ages, sex, aborted and pregnant, and lack of controlled movement of animals. Therefore, effective control of animal brucellosis requires surveillance to identify infected animal herds, elimination of the reservoirs, and vaccination of young heifers. B. melitensis biovar 3 is the cause of the Brucella outbreak in buffalo which still remains the prevalent type of Brucella in Egypt. The disease runs a chronic course allowing further spread of infection. Keywords: bruce-ladder, brucellosis, buffalo, histopathology, polymerase chain reaction.

The aim of this study was to investigate the effects of phytase which was laboratory produced by Aspergillus foetidus on the growth performance, mineral retention, and bone traits of broilers fed with low dietary calcium and phosphorus. The extracellular phytase enzyme secreted into the crude filtrate was concentrated by ammonium sulfate precipitation to obtain an activity of 500 phytase units (FTU). A total of 90 1-day-old chicks (Cobb 500) were randomly divided into three treatment groups with five replicates having six birds each. Dietary treatment, T1, was with 0.45% non-phytate P (NPP) during starter and 0.40% during finisher phase with 1% Ca. Dietary treatment, T2, had 0.37% NPP during starter and 0.32% in finisher phase with 1% Ca and supplemental lab phytase at 500 FTU/kg. Dietary treatment, T3, was similar to T2 with a lower Ca of 0.8%. There was no significant difference among the dietary treatments with regard to body weight gain, feed intake, feed conversion ratio, and Ca retention (p>0.05). However, a significant improvement in retention of P by birds was observed in phytase supplemental groups T2 and T3 (p<0.05). Dry weight of tibia (2.58-2.78 g/kg live weight) and ash content (39.7- 41.8%) was comparable among treatments. A similar trend was observed for bone Ca, P, and Mn content. The study indicated that 500 FTU/kg phytase can be effectively supplemented in a broiler diet with low phosphorus (0.37% in starter and 0.32% NPP in finisher diet) and low calcium (0.8% in diet) for better growth performance and with successful replacement of dietary P by 0.08 % and reduced P excretion into the environment in broiler chicken. Keywords: broiler, calcium, phosphorus, phytase.

The immunocompromised condition is considered a defect in the immune system. This condition tends to increase the risk of oral candidiasis, due to the inability of the immune system to eliminate the adhesion of Candida albicans and leads to systemic candidiasis with a mortality rate of 60%. Green tea (Camellia sinensis) contains potential antioxidant and immunomodulatory which acts as anticancer, antifungal, and antivirus agent. The aim of this study was to invent herbal-based medicine, which acts as an immunomodulator and antifungal agent to treat fungal infection in immunocompromised patients. Thirty-five immunocompromised Wistar rats induced with C. albicans were divided into 7 groups (n=5): Control group (C+); treated for 4 days with green tea extract 1.25% (GT 4), epigallocatechin gallate (EGCG) 1% (EGCG 4), EGC 1% (EGC 4); and treated for 7 days with green tea extract 1.25% (GT 7), EGCG 1% (EGCG 7), and EGC 1% (EGC 7). Tongue tissue was collected and analyzed with immunohistochemistry staining using monoclonal antibody; interleukin (IL)-17A, IL-8, and human beta-defensin 2 (HBD)-2. Data were analyzed using analysis of variance test and Tukey honest significant differences test. The expression of IL-17A, IL-8, and HBD-2 was significantly increased (p=0.000) after green tea extract administration in 7 days, whereas in 7 days, the expression of IL-8, IL-17A, and HBD-2 after EGCG and EGC administration did not give a significant result (p>0.005). Within the limits of this study, green tea extract has the ability as an immunomodulatory agent in an immunocompromised patient infected by C. albicans through expression augmentation of IL-8, IL-17A, and HBD-2 compared to EGCG and EGC. Keywords: epigallocatechin gallate, epigallocatechin, green tea extract, immunocompromised, oral candidiasis.

The current study aimed to compare the impact of heat stress (HS) on some physiological functions and blood oxidative stress biomarkers between dry dairy crossbred (Balady X Friesian) cows and buffaloes during Egyptian summer season (July-September). A total of 26 healthy animals were equally used in the in the current study. The criterion for cows and buffaloes selection and the management conditions were similar. A total mixed ration to meet the animal's requirements was used, and dry matter intake (DMI) was calculated. Ambient temperature, relative humidity, temperature humidity index (THI), respiratory rate, and rectal temperature (RT) were daily recorded. Meanwhile, live body weight and body condition score were weekly recorded. Blood samples were collected bi-weekly, and plasma samples were harvested for malondialdehyde (MDA) content and enzymatic antioxidants such as glutathione peroxidase, superoxide dismutase, and catalase activities determinations throughout the experimental period (8 weeks - prepartum). The results confirmed, the HS condition, as the THI values ranged from 79.74 to 90.4 throughout the experimental period. In both species, HS increased RT and decreased DMI (<10.5 kg/day and 9.5 kg/day in cows and buffaloes, respectively). Buffaloes seemed to be more affected by the hostile environmental condition of this study compared with their respective cows. Buffaloes had recorded up to 1 °C increase in their RTs in most of the point's period compared to cows. There was a continuous increase in MDA values (194.7 and 208.4 nmol/gHb in buffaloes and cows, respectively, 2 weeks prepartum) as the animals come close to parturition with moderate decrements for the enzymatic antioxidant activities in both cows and buffaloes. It can be concluded that during Egyptian's summer season, HS had adversely affected feed intake and consequently animal's production performances. Keywords: buffaloes, dairy cows, Egyptian's summer, heat stress, oxidative stress.

This study was conducted to study the genetic and population structure of local (Awassi) and exotic (Romanov, Charollais, Assaf, and Suffolk) sheep breeds in Jordan using eight microsatellite markers. A total of 125 sheep were used (25 from each breed) in the study. The number of alleles (A), the mean values of observed (Ho) and expected (He) heterozygosity, polymorphism information content (PIC), fixation index as a measure of heterozygote deficiency or excess, and Hardy-Weinberg equilibrium (HWE) were analyzed using PopGen and CERVUS softwares. Nei's standard genetic distances among breeds and dendrogram of unweighted pair group method with arithmetic mean (UPGMA) were calculated and constructed using PopGen software. A total of 40 alleles were detected with an average number of alleles of 5. The mean Ho value was higher than the mean He value for all breeds. Awassi breed showed the highest average PIC value while Romanov had the lowest. There was a significant (p<0.05) deviation from HWE at each locus within and between breeds. Deviations from HWE were found to be highly significant for all markers except OARFCP304 locus. The genetic distance estimates revealed a close relationship between Romanov and Charollais and between Awassi and Charollais. In the UPGMA dendrogram, Charollais, Romanov, and Awassi breeds were placed together in one main cluster while Assaf was in a different subcluster. Awassi was placed alone in a second main cluster. Results of this study offer insight toward the genetic conservation of the studied breeds and a base on which breeding plans can be made. Keywords: genetic diversity, Jordan, microsatellites, sheep breeds.

We examined regulatory function of astaxanthin on mRNA expression of nuclear factor κB (NF-κB) p65, interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), and interferon gamma (IFN-γ) in peripheral blood mononuclear cells in pre and postpartum Murrah buffaloes during summer (temperature-humidity index [THI]=86; relative humidity [RH]=24) and winter (THI=58.74; RH=73) seasons. A total of 32 Murrah buffaloes apparently healthy and in their one to four parity were selected from National Dairy Research Institute herd and equally distributed randomly into four groups (control and supplemented groups of buffaloes during summer and winter season, respectively). All groups were fed according to the nutrient requirement of buffaloes (ICAR, 2013). The treatment group was supplemented with astaxanthin at 0.25 mg/kg body weight/animal/day during the period 30 days before expected date of calving and up to 30 days postpartum. There was downregulation of NF-κB p65 gene in all the groups. NF-κB p65 mRNA expression was lower (p<0.05) in treatment than control group from prepartum to postpartum during summer, while mRNA expression was low only on day 21 after calving during winter season. The mRNA expression of IL-6, TNF-α, and IFN-γ was lower (p<0.05) in treatment than a control group of buffaloes during summer and winter seasons. The mRNA expression of NF-κB p65, IL-6, TNF-α, and IFN-γ was higher (p<0.05) in summer than in winter seasons. The xanthophyll carotenoid astaxanthin a reddish-colored C-40 compound is a powerful broad-ranging antioxidant that naturally occurs in a wide variety of living organisms, such as microalgae, fungi, crustaceans, and complex plants. Astaxanthin blocked nuclear translocation of NF-κB p65 subunit and IκBa degradation, which correlated with its inhibitory effect on IκB kinase (IKK) activity. These results suggest that astaxanthin, probably due to its antioxidant activity, inhibits the production of inflammatory mediators by blocking NF-κB activation and as a consequent suppression of IKK activity and IκB-degradation. Keywords: astaxanthin, Murrah buffalo, nuclear factor κB p65, summer, winter.

Somatic cell count (SCC) is the most widely used single reliable indicator of udder health. The present study was carried out with an objective to find the exact threshold of SCC. Milk samples collected from a total of 214 Holstein Friesian crossbred dairy animals were subjected to bacterial DNA extraction and SCC estimation by digital PortaCheck. California Mastitis Test and polymerase chain reaction based on amplification of organism using reported primers were performed to diagnose subclinical mastitis. Receiver's operating characteristic (ROC) curve analysis and discriminate function analyses were performed using SPSS 18 software. ROC curve analysis represented that the area under the curve was 0.930 with the standard error of 0.02. Results indicated that 93% of the case could be correctly predicted as mastitis infected using SCC as a marker (p<0.001). At cut score level of 282 000 cells/ml, 285,000 cells/ml and 288,000 cells/ml, sensitivity remained 92.6% and specificity augmented as 86.3%, 87.2%, and 88%, respectively. At SCC value of 310,000 cells/ml of milk, sensitivity and specificity were optimal, namely, 92.6% and 91.5%, respectively. The function fitted demonstrated 89.2% accuracy with p<0.001. The functions at group centroids were -0.982 and 1.209, respectively, for normal and mastitis-infected animals and log_SCC value was the most important factor contributing 38.30% of the total distance measured. Our study supports that the threshold value to delineate subclinical mastitis case from the normal is 310,000 somatic cells/ml of milk and a model so fitted using the variable SCC can be successfully used in field for the diagnosis of subclinical cases of mastitis which otherwise would be difficult to differentiate based on clinical signs. Keywords: discriminate function, mastitis, receiver's operating characteristic curve, somatic cell count, threshold.

The major objective of the investigation was to evaluate the hitherto uncharacterized potential of Brucella-specific antibodies to win the battle against virulent Brucella abortus infection. Brucella-specific immune serum was raised in mice. The antibody titer of serum was determined by standard tube agglutination test and indirect enzyme-linked immunosorbent assays (iELISA). Groups of mice and guinea pigs were passively immunized with serum containing specific agglutinin titers. 24 h after immunization, all animals along with unimmunized controls were challenged with B. abortus S544. Total B. abortus S544 counts in the spleen of each animal collected on the 7th day of challenge was determined to evaluate the protective index (PI) of anti-Brucella serum by statistical analysis. A dose-dependent protective response to immune mice serum was observed in both experimental models though the values of PI of mice were higher than those obtained for guinea pigs. The PI values in mice passively immunized with 50 IU or 25 IU antibodies were 1.38 and 0.69, respectively. In guinea pigs, however, animals passively immunized with 50 IU or 25 IU antibodies showed PI values equivalent to 0.79 and 0.41, respectively. The observations support our hypothesis that the presence of antibodies inhibits the initial multiplication and eventual colonization of systemic organs by B. abortus. Therefore, a predominant antibody-mediated response induced by a vaccine is expected to protect the animal against the most severe clinical outcome of infection. Keywords: brucellosis, humoral immunity, mice, passive protection, protective index.

The therapeutic evaluation of the biological effect of platelet-rich plasma (PRP) used as a surgical adjunct to maintain the inflammatory process and to potentiate tissue healing, make the subject of recent research in regenerative medicine. This study was designed to evaluate the healing activity of PRP by its topical application on the skin experimentally injured in a sheep model. The study was conducted on 9 adult and clinically healthy males sheep. PRP was obtained by a protocol of double centrifugation of whole blood from each animal. After sterile skin preparation, full-thickness excisional wounds (20 mm x 20 mm) were created on the back of each animal. The animals were randomly divided into three equal groups of three sheep for each. In Group I, the wounds were treated with PRP, in Group II; wounds were treated with Asiaticoside; in Group III, wounds were treated with saline solution. The different treatments were administered topically every 3 days. Morphometric measurements of the contraction surface of the wounds and histopathological biopsies were carried out at the 3rd, 7th, 14th, 21st, and 28th days of healing. The results of the morphometric data obtained revealed that it was significant differences recorded at the 7th and 14th day of healing in favor for animals of Group I. Semi-quantitative histopathological evaluation showed that PRP reduces inflammation during 3 first days post-surgical and promotes epithelialization in 3 weeks of healing. We concluded that topical administration of PRP obtained by double centrifugation protocol could potentially improve the skin healing process in sheep. Keywords: healing, platelet-rich plasma, sheep, skin.

The aim of the study was to explore the anti-methanogenic potential of phyto-sources from Northeast region of the country and assess the effect on rumen fermentation characteristics and protozoa for their likely inclusion in animal diet to reduce methane emission. Twenty phyto-sources were collected from Northeast state, Assam, during March to April 2014. Phyto-sources were analyzed for their tannin content followed by screening for methane mitigation potential using in vitro system. The effect of tannin on methane production and other fermentation parameters was confirmed by attenuating the effect of tannin with polyethylene glycol (PEG)-6000 addition. About 200 mg dried phyto-source samples were incubated for 24 h in vitro, and volume of gas produced was recorded. The gas sample was analyzed on gas chromatograph for the proportion of methane in the sample. The effect of phyto-sources on rumen fermentation characteristics and protozoal population was determined using standard methodologies. Results from studies demonstrated that Litchi chinensis, Melastoma malabathricum, Lagerstroemia speciosa, Terminalia chebula, and Syzygium cumini produced comparatively less methane, while Christella parasitica, Leucas linifolia, Citrus grandis, and Aquilaria malaccensis produced relatively more methane during in vitro incubation. An increase (p<0.05) in gas and methane production from the phyto-sources was observed when incubated with PEG-6000. Entodinimorphs were prominent ciliates irrespective of the phyto-sources, while holotrichs represented only small fraction of protozoa. An increase (p<0.05) in total protozoa, entodinimorphs, and holotrichs was noted when PEG-6000 added to the basal substrate. Our study confirmed variable impact of phyto-sources on total volatile fatty acid production and ammonia-N. It may be concluded that L. chinensis, M. malabathricum, L. speciosa, S. cumini, and T. chebula are having potent methane suppressing properties as observed in vitro in 24 h. These leaves could be supplemented in the animal diet for reducing methane emission; however, in vivo trials are warranted to confirm the methane inhibitory action and optimize the level of supplementation. Keywords: methane, phyto-sources, protozoa, rumen fermentation, tannin.

Scabies is one of the most important diseases in goats and caused by a complex hypersensitivity process that involves both humoral and cell-mediated immune responses. This phenomenon shows that the variety of Sarcoptes scabiei has different characteristics of specific antigenic protein or different immune-dominant. This research aims to detect the humoral and cellular immune response of rabbits which were immunized with the protein of S. scabiei var. caprae. This research was done as follows, identification and collection of Sarcoptes scabiei var. caprae from goat infected with scabies, separation of protein antigen from S. scabiei mites with ultrasonic sonicator, measurement of protein content with spectrophotometry, rabbit injection with 500 μg dose of antigen protein which was repeated 5 times (5x booster) every 2 weeks. Measurement of IgG titer using indirect ELISA, whereas to detect the expression of cellular immune response (TLR-9, CD4, and CD8) using Direct Immunofluorescence assay. Based on the statistical analysis, it showed that there was a significant enhancement (p<0.05) for optical density value or antibody titer and cellular immune response was shown by TLR-9, CD4, and CD8 expression in rabbit T lymphocytes which appear yellow to green fluorescent color using fluorescence microscope. The amount of fluorescence T lymphocytes showed a significant difference (p<0.05) between the control and various boosters. Antigenic protein of S. scabiei var. caprae contains ligands, which are involved in the pathogen-associated molecular pattern that has an ability to induce humoral and cellular immune response in rabbit. Specifically, that TLR-9 is not only involved in innate immunity but also in adaptive immunity and can be used as alternative adjuvant development research. Keywords: antigenic protein, CD4, CD8, immunoglobulin G, Sarcoptes scabiei var. caprae, toll-like receptor-9.

Investigation of antiviral activity of Acanthaster planci phospholipase A2 (AP-PLA2) from moluccas to human immunodeficiency virus (HIV). Crude venom (CV) and F20 (PLA2 with 20% fractioned by ammonium sulfate) as a sample of PLA 2 obtained from A. planci's extract were used. Enzymatic activity of PLA2 was determined using the degradation of phosphatidylcholine (PC). Activity test was performed using in vitro method using coculture of phytohemagglutinin-stimulated peripheral blood mononuclear cell (PBMC) from a blood donor and PBMC from HIV patient. Toxicity test of AP-PLA2 was done using lethal concentration required to kill 50% of the population (LC50). AP-PLA2 F20 had activity and purity by 15.66 times bigger than CV. The test showed that the LC50 of AP-PLA2 is 1.638 mg/ml. Antiviral analysis of AP-PLA2 in vitro showed the inhibition of HIV infection to PBMC. HIV culture with AP-PLA2 and without AP-PLA2 has shown the number of infected PBMC (0.299±0.212% and 9.718±0.802%). Subsequently, RNA amplification of HIV using reverse transcriptase-polymerase chain reaction resulted in the decrease of band intensity in gag gene of HIV. This research suggests that AP-PLA2 has the potential to develop as an antiviral agent because in vitro experiment showed its ability to decrease HIV infection in PBMC and the number of HIV ribonucleic acid in culture. Keywords: Acanthaster planci, antiviral activity, human immunodeficiency virus, Indonesia, phospholipase.

Milk adulteration is pivotal because it leads to worse effects in public health as human adverse reactions with clinical signs ranged from gastrointestinal signs to anaphylactic shock. This study was carried out to estimate the prevalence of adulteration in buffalo's milk sold in Assiut City, Egypt. A total of 50 raw buffalo's milk samples were collected and examined for adulteration by addition of cow's milk. The examination carried out by applying polymerase chain reaction-restriction fragment length polymorphism technique using cytochrome b (cyt b) gene primers and Hinf I enzymes. The size of target gene was 360 bp in both animal species and amplicon can be digested using Hinf I enzyme, this restriction enzyme divided the essential band to clear three bands at 360, 210, and 150 bp in cows' milk, while, the enzyme could not be cleaved the amplicon in buffalo's samples. The obtained results cleared that the incidence of adulteration of buffalo's milk very high percentage reaches 90%. It could be concluded that the raw buffalo's milk sold in Assiut City subject to fraudulent practice and thus can lead to public health hazards. Keywords: buffalo's milk, cyt b gene, Hinf I enzyme, medicolegal, milk adulteration, polymerase chain reaction-restriction fragment length polymorphism.

This study aimed to evaluate the effects of Meniran extract (Phyllanthus niruri L.) administration on leukocyte profile of broiler chickens infected with Mycoplasma gallisepticum. Thirty broiler chickens, 21 days old were divided into five treatment groups. P0 (-): Chickens without any treatment; P0 (+), P1, P2, and P3: Chickens were infected with M. gallisepticum 108 cells/ml/animal orally, then given no treatment, Meniran extract 60%, 62.5%, and 65% orally at a dose of 1 ml/kg body weight, respectively. The treatment of Meniran extract was given for 7 days. Leukocyte count with the lowest number showed in Group P0 (-) and Group P3 (p>0.05). Increased number of basophils was found in Group P0 (+), Group P1, and Group P2. The highest number of heterophils was found in Group P0 (+) and was significantly different from Group P0 to P3 (p<0.05). The same pattern was also seen in the number of lymphocytes in all treatment groups. The number of monocytes showed no significant difference between all treatment groups (p>0.05). Discussion: Increased the number of leukocytes is often observed in inflammation due to general infections, trauma, or toxicity. Shifting in the number of heterophile or lymphocytes, an increase in the number of monocytes, basophils, and eosinophils may also be associated with various infectious or inflammatory conditions. Heterophils play a role as an antibacterial defense through several effective mechanisms. When infections and inflammation occur, the heterophils will increase to phagocytosis microbe. It can be concluded that Meniran extract (P. niruri L.) at a dose of 65% can decrease the total number of leukocytes in broilers infected with M. gallisepticum. Keywords: chicken, leukocyte, Mycoplasma gallisepticum, Phyllanthus niruri L.

The objectives of this study were to determine the leptospirosis seroprevalence and to identify the predominant infecting serovars among cattle. A cross-sectional study involving 420 cattle from six randomly selected districts in Kelantan was conducted. A serological test using the microscopic agglutination test was conducted in the Institute of Medical Research with a cutoff titer for seropositivity of ≥1:100. The overall prevalence of leptospirosis seropositivity among cattle in this study was 81.7% (95% confidence interval: 63.5, 80.1). The most common reaction obtained with the sera tested was from the serovar Sarawak with 78.8%. A high seroprevalence of leptospiral antibodies was found among cattle in Northeastern Malaysia. These findings urge that more studies are required to determine the reasons for the high seroprevalence among the cattle along with its transmission and pathogenicity of the local serovar Sarawak. Keywords: cattle, leptospirosis, microscopic agglutination test, serovar Sarawak.

Rhipicephalus microplus (Rm) is one of the most problematic livestock tick species in the world. Its rapid propagation and resistance to acaricides make it control difficult in the sub-region and Benin particularly. The aim of this work was to check its presence in wildlife and to confirm the possible role of reservoir wildlife may play in the propagation of the parasite. This will help to design more efficient control strategy. This study was conducted from February to March 2017 in the National Parks of Benin (Pendjari and W Park) and wildfowl's assembly and selling point in Benin. Ticks were manually picked with forceps from each animal after slaughtering by hunters then stored in 70° ethanol. Collected ticks were counted and identified in the laboratory using the identification key as described by Walker. Overall, seven species of ticks (Amblyomma variegatum, Boophilus decoloratus, Rm, Boophilus spp., Hyalomma spp., Rhipicephalus sanguineus, Rhipicephalus spp.) were identified on nine wild animal species sampled (Cane rat, wildcat, Hare, Doe, Cricetoma, Buffalo, Buffon Cobe, and Bushbuck and Warthog). The average number of ticks varies from 3 to 6 between animal species, 3 to 7 between localities visited, and 2 to 5 between tick species. However, these differences are statistically significant only for localities. Considering tick species and animal species, the parasite load of Rm and Rhipicephalus spp. is higher; the buffalo being more infested. The analysis of deviance reveals that the abundance of ticks observed depends only on the observed localities (p>0.05). However, the interactions between animal species and localities on the one hand and between animal and tick species on the other hand, although not significant, have influenced the abundance of ticks as they reduce the residual deviance after their inclusion in the model. This study reported the presence of Rm in wildlife of Benin and confirmed its role in the maintenance and spread of the parasites. It is, therefore, an important risk factor that we must not neglect in the epidemiological surveillance and ticks control strategies in the West African sub-region and particularly in Benin. Keywords: Benin, Rhipicephalus microplus, ticks, wild animals.

Regulation of pH in spermatozoa is a complex and dynamic process as sperm cells encounter different pH gradients during their journey from testes to the site of fertilization in female genital tract. The precise regulations of pH in sperm cells regulate the sperm functions such as motility, hyperactivity, capacitation, and acrosome reaction. Electrophysiological, pharmacological, and molecular studies have revealed the presence of different ion channels and exchanger systems which regulate intracellular pH in sperm cells as well as regulate sperm functions. Recent studies also have shown the potential involvement of pH in the regulation of fertility competence of sperm cells, and alterations in pH have shown to impede sperm functions. This mini-review discusses the probable mechanisms involved in pH regulation in sperm cells and how pH is involved in regulation of various sperm functions.

Avian fecal Escherichia coli (AFEC) are considered to be the natural reservoir of pathogenic strains in extraintestinal infections as such characterization of AFEC gives insight into the spread of the potential pathogenic lineage. The aim of the study was to investigate the reservoirs of avian pathogenic E. coli (APEC) from fecal samples of healthy ducks, geese, and turkeys by determining the antibiotic resistance patterns of AFEC isolates from turkeys, geese and ducks and characterization of the isolates using virulence genes, plasmid profiles, and phylogenetic grouping. The disc diffusion method was used to determine antibiotic resistance of 100 AFEC isolates from turkeys (9), geese (29), and ducks (62) to 8 antibiotics. Molecular characterization of the isolates was done by multiplex polymerase chain reaction to investigate the presence of 12 virulence genes, plasmid profiling, and phylogenetic grouping based on the 16S rRNA sequences. Antibiogram profiles indicated maximum resistance to cloxacillin (100%) and bacitracin (100%) for all AFEC isolates and high sensitivity to ciprofloxacin; however, all isolates exhibited multi-drug resistance. The AFEC isolates from turkeys (6) and geese (12) did not contain virulence genes. The frz (3.7%), sitD (29.6%), and fimH (92.5%) were detected in the duck isolates. None of the isolates had the KpsM, iutA, vat, sitA, hlyF, pstB, ompT, uvrY, and sopB genes. Plasmid profiling gave four plasmid profiles with the plasmids ranging from 1.5 to 55 kb. Phylogenetic analysis of 16S rRNA sequences revealed similarities between AFEC isolates from the different poultry species, as the isolates did not cluster according to avian species. AFEC isolates are potential reservoirs of APEC as they contain some of the virulence genes associated with APEC. Multidrug resistance is high in AFEC isolated from healthy birds. This is a public health concern. Keywords: antibiotic resistance, avian fecal Escherichia coli, poultry, virulence gene.

The present study was conducted to record the prevalence of extended spectrum β-lactamases (ESBLs) producing Escherichia coli, Salmonella spp., and Klebsiella pneumoniae from pig population of Assam and Meghalaya and to record the ability of the resistant bacteria to transfer the resistance genes horizontally. Fecal samples (n=228), collected from pigs of Assam (n=99) and Meghalaya (n=129), were processed for isolation and identification of E. coli and Salmonella spp. All the isolates were tested for ESBLs production by double disc synergy test (DDST) followed by screening for ESBLs producing genes (blaTEM, blaSHV, blaCTX-M, and blaCMY) by polymerase chain reaction (PCR). Possible transfer of resistance encoding genes between enteric bacterial species was carried out by in vitro and in vivo horizontal gene transfer (HGT) method. A total of 897 enteric bacteria (867 E. coli and 30 Salmonella) were isolated and identified. Altogether 25.41% isolates were confirmed as ESBL producers by DDST method. Majority of the isolates were E. coli followed by Salmonella. By PCR, 9.03% isolates were found positive for at least one of the target resistance genes. blaSHV was absent in all the isolates. blaCMY was the most prevalent gene. All the E. coli isolates from Assam were negative for blaTEM. A total of 2.76% isolates were positive for blaTEM + blaCMY. On the other hand, 0.67% isolates were positive for blaCTX-M + blaCMY genes. Only 0.33% isolates carried all the three genes. Altogether, 4.68% bacteria carried the resistance encoding genes in their plasmids. blaTEM gene could be successfully transferred from Salmonella (donor) to E. coli (recipient) by in vitro (5.5-5.7x10-5) and in vivo (6.5x10-5 to 8.8x10-4) methods. In vivo method was more effective than in vitro in the transfer of resistance genes. The pig population of Assam and Meghalaya are carrying multidrug resistance and ESBLs producing E. coli and Salmonella. The isolates are also capable to transfer their resistance trait to other bacterial species by HGT. The present finding could be considered as a serious public health concern as similar trait can also be transmitted to the human commensal bacteria as well as pathogens. Keywords: Enterobacteriaceae, multidrug resistance, North East India, pigs.

The aim of this study was to investigate the effect of dietary supplementation with multi-strain probiotic preparation in combination with vitamins and minerals on the hematological parameters and selected intestinal microbiota populations in the Indonesian indigenous crossbred chickens. A total of 240 one-day-old Indonesian indigenous crossbred chicks were raised for 10 weeks. The chicks were distributed to one of four groups, i.e., chicks receiving basal diet without any additive (CONT), chicks receiving basal diet with 0.04% of zinc bacitracin (AGP), chicks receiving basal diet with 0.01% of commercial probiotic Bacillus subtilis preparation (PROB1), and chicks receiving basal diet with 0.5% of multi-strain probiotic preparation in combination with vitamins and minerals (PROB2). Blood was collected on the week 8, while the internal organs and eviscerated carcasses were collected on the week 10. PROB2 tended (p=0.09) to have a lower body weight (BW) compared to CONT chicks. Feed conversion ratio was higher (p<0.05) in PROB1 and PROB2 compared to CONT birds. The number of thrombocytes tended (p=0.09) to be higher in CONT than in other groups. Antibody titer against Newcastle disease virus vaccine was higher (p<0.05) in PROB1 and PROB2 than in CONT group. Serum triglyceride concentration was lower (p<0.05) in PROB2 than in other birds. AGP chicks had lower (p<0.05) serum total protein and globulin concentrations than CONT and PROB1 chicks. Serum albumin level was lower (p<0.05) in PROB2 than in CONT and PROB1 birds. Albumin to globulin ratio tended (p=0.06) to be higher in AGP than in other birds. Lactose-negative Enterobacteriaceae tended (p=0.07) to be lower in PROB1 and PROB2 than in CONT group. PROB1 and PROB2 tended (p=0.06) to have greater lactic acid bacteria (LAB) population than in CONT and AGP birds. Multi-strain probiotic preparation in combination with vitamins and minerals was able to improve immune response and control the potentially pathogenic bacteria. However, the additive could not improve the growth performance of the Indonesian indigenous crossbred chickens. Keywords: growth performance, Indonesian indigenous crossbred chickens, intestinal bacteria, minerals, multi-strain probiotic, vitamins.

During the last decades, reproduction performances declined dramatically worldwide, but little is known concerning the involvement of oxidative stress as a causative factor. Oxidative stress may act at different levels, with negative impacts on cell membrane integrity and other active molecules with potential subsequent effects on reproduction. The aim of the current study was to investigate the oxidative stress status in cows according to their reproductive performances. Peripheral blood concentration of two oxidative stress biomarkers, glutathione S-transferase (GST) and malondialdehyde (MDA), and other biochemical parameters (glucose, total lipids, cholesterol, triglycerides, albumin, total proteins, calcium, urea, creatinine, direct bilirubin, alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase) were determined in 40 healthy cows. Body condition score (BCS), calving to first service interval (FSI), calving to conception interval (CCI), and the number of service per conception (SPC) were simultaneously recorded for each cow. Concerning FSI, three groups were established: Group 1 (from 44 to 60 days), Group 2 (from 60 to 70 days), and Group 3 (from 70 to 80 days). For CCI, two groups were considered: Group 1 (<110 days) and Group 2 (>110 days). MDA showed significant high values only in cows with the lowest BCS (1.5) compared to cows with BCS note of 2.5 and 3.5. No significant difference was observed in cows oxidative stress status (MDA and GST) according to reproductive performances (FSI, CCI, and SPC) in all studied groups. The results revealed relatively altered oxidative stress status in cows with abnormal reproductive performances; however, no significant difference was recorded whatever the considered reproductive parameter. Keywords: cow, glutathione S-transferase, malondialdehyde, reproduction performance.