Antimicrobial resistance is an emerging public health threat. Foodborne illnesses are typically caused by bacteria, such as Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, and Staphylococcus aureus, which are frequently resistant to common antimicrobial agents. Rice is a staple grain in most parts of the world. Our previous work showed that Phatthalung Sangyod rice seed protein hydrolysates (SYPs), especially SYP4, exhibit antifungal activity against several fungal species that are pathogenic for both humans and animals and are non-cytotoxic to animal red blood cells. In this study, we aimed to determine the effects of the bioactive peptides in SYPs against several pathogenic bacteria in humans and animals. After isolating SYP1, it was treated as follows: heated (SYP2), and hydrolyzed using pepsin (SYP3), and proteinase K (SYP4). Then, we used 500 μg of protein to evaluate the antibacterial effects on four pathogenic bacteria, including E. coli, P. aeruginosa, B. cereus, and S. aureus, using agar well diffusion. Using a broth microdilution assay, we determined the minimum inhibitory and bactericidal concentration (MIC and MBC, respectively) values of active SYPs. Using the agar well diffusion and microtube incubation methods, we also assessed the inhibitory effects of SYPs on the bacterial quorum sensing (QS) activity of Chromobacterium violaceum. Sangyod rice seed protein hydrolysates were evaluated for their ability to inhibit the biofilm formation of bacterial cells by a crytal violet assay. Furthermore, using the dropping method, we tested the inhibitory effects of SYPs on the bacterial pigments pyocyanin in P. aeruginosa and staphyloxanthin in S. aureus. Our results showed that the crude protein lysate (SYP1) did not exhibit antibacterial activity against any of the test bacteria. Intriguingly, after boiling (SYP2) and enzymatic hydrolysis (SYP3 and SYP4), the protein hydrolysates were transformed into bioactive peptides and displayed antibacterial properties against all of the test bacteria at a concentration of 500 μg as determined by agar well diffusion. SYP4 demonstrated the highest antibacterial activity as it completely inhibited all test strains, with inhibition zones ranging from 16.88 ± 0.25 to 21.25 ± 0.5 mm, and also yielded the highest MIC/MBC values against P. aeruginosa, B. cereus, and E. coli, at 256 and >256 μg/mL, respectively. We observed that at least 256 μg/mL of SYP4 is required to exhibit optimal antibacterial activity. At 16–128 μg/mL, it exhibited antibiofilm activity against S. aureus. Furthermore, at 256 μg/mL, SYP4 inhibited pyocyanin in P. aeruginosa and staphyloxanthin in S. aureus. Although SYP2 and SYP3 displayed weak antibacterial activity and their MIC values could not be obtained for all bacteria, they showed strong QS inhibition in C. violaceum at 256 μg protein. Moreover, SYP2 and SYP3, at a minimum concentration of 32 μg/mL, significantly reduced violacein production. SYP3 also showed biofilm reduction activity on S. aureus at least 16-512 μg/mL. Sangyod Phatthalung protein hydrolysates exerted excellent inhibitory effects against the growth of bacteria and their virulence factors, such as QS, biofilm formation, and/or pigment production. These factors include zoonotic and foodborne pathogens. Therefore, daily consumption of Sangyod Phatthalung rice might reduce the risk of bacterial pathogenesis and foodborne diseases. In conclusion, functional foods or alternate methods of treating bacterial illnesses may be developed in humans and animals. Keywords: antibacterial peptide, foodborne pathogens, Phatthalung Sangyod rice, protein hydrolysate, zoonotic.

Foodborne diseases caused by Escherichia coli are prevalent globally. Treatment is challenging due to antibiotic resistance in bacteria, except for foodborne infections due to Shiga toxin-producing E. coli, for which treatment is symptomatic. Several studies have been conducted in Africa on antibiotic resistance of E. coli isolated from several sources. The prevalence and distribution of resistant pathogenic E. coli isolated from food, human, and animal sources and environmental samples and their virulence gene profiles were systematically reviewed. Bibliographic searches were performed using four databases. Research articles published between 2000 and 2022 on antibiotic susceptibility and virulence gene profile of E. coli isolated from food and other sources were selected. In total, 64 articles were selected from 14 African countries: 45% of the studies were conducted on food, 34% on animal samples, 21% on human disease surveillance, and 13% on environmental samples. According to these studies, E. coli is resistant to ~50 antimicrobial agents, multidrug-resistant, and can transmit at least 37 types of virulence genes. Polymerase chain reaction was used to characterize E. coli and determine virulence genes. A significant variation in epidemiological data was noticed within countries, authors, and sources (settings). These results can be used as an updated database for monitoring E. coli resistance in Africa. More studies using state-of-the-art equipment are needed to determine all resistance and virulence genes in pathogenic E. coli isolated in Africa. Keywords: Africa, antibiotic resistance, Escherichia coli virulence genes, food, systematic review.

Poultry production is the fastest growing livestock industry in the world, as the rapid growth of and efficient absorption of feed by poultry ensure the production of poultry meat with a relatively low carbon footprint. Seeking new ways to increase livestock productivity as well as poultry product quality, the number of research studies on the use of humic substances of various origins in livestock farming has increased significantly, emphasizing the role of feed additives derived from local resources. The unique capability of humic substances to improve metabolic processes allows the immune protection of the bird body to be strengthened and production efficiency to be increased. This study aimed to identify the effects of sodium humate (NaHum) on the growth performance of broiler chickens and selected blood and ileum microbiota parameters. Dietary research was conducted 2 times under production conditions in a poultry facility of a commercial company, with 210 1-day-old, unsorted broiler chickens of both sexes (Ross 308). The broiler chickens were fed with standard commercial feed, the rearing period of 35 days, and slaughtered on day 36. Sodium humate additive was added to drinking water for the research groups of broilers in period from 8th to 35th day of life, 25 mL (Group 1, n = 2 × 35) and 50 mL (Group 2, n = 2 × 35) per liter of drinking water. Sodium humate contained an average of 4.48% dry matter, a kilogram of dry matter containing 104.3 g of crude protein, 3.6 g of crude fiber and 0.9 g of crude fat, 14.3 MJ of metabolic energy, and 5.8 MJ of energy for live weight gain, as well as a very high content of crude ash −759.8 g, including 4.2 g Ca, 4.2 g Na, and 4.81 g Fe, the dry matter digestibility of NaHum was 87.0%, and the absorption capacity of dry matter was 113.2%., the pH level was 13.0, i.e., alkaline. At the end of the dietary research, the productivity and economic efficiency of the research groups of broilers were calculated by live weight gain, carcass weight, feed conversation ratio, and blood and intestinal samples of broilers were analyzed to identify the effects of NaHum on the growth performance and health status of broilers. Dietary research found that adding 25 mL/L and 50 mL/L of NaHum to drinking water for the broiler chickens increased their live weights at the selling age, average live weight gains by 3.06–3.93%, and carcass weights by 5.07–6.06%, while feed conversion increased in terms of both live weight (1.5 and 1.51) and carcass weight (1.84 and 1.86) compared with the control group. The best economic performance in terms of the economic efficiency index and the cost index (CI) was found in Group 1, which was fed with the NaHum additive at an intake rate of 25 mL/L. The NaHum additive modulated the ileal microbiota and metabolic processes in the broiler body. At the same time, a significant decrease in the levels of total protein, alkaline phosphatase and phosphorus (P) in blood was found in the research groups. Considering the positive effects of NaHum derived from freshwater sapropel on the productivity and economic efficiency of broiler chickens, the NaHum feed additive should be further investigated on a larger scale to obtain results that could reasonably be used in practice. This study concluded that a decrease in P levels in the blood was observed when NaHum was added to the drinking water; therefore, it is important to continue the research to draw reasonable conclusions on the effects of NaHum in liquid form on the health performance of farm animals. Keywords: broilers, carcas yield, growth performance, ileum microbiota, sapropel, sodium humate.

Canine babesiosis, caused by the protozoan parasite Babesia canis, is characterized by clinical manifestations, including hemolytic anemia, thrombocytopenia, multiple organ failure, and may result in death. This disease is detected using conventional blood smears, which are time-consuming and have low sensitivity. This study aimed to investigate a more rapid and sensitive method for detecting B. canis infection in dogs by examining the expressed serum protein profiles using proteomics. We collected six sera samples from three healthy and three B. canis-infected dogs diagnosed using blood smear and polymerase chain reaction. We analyzed the proteins using two-dimensional gel electrophoresis. The candidate spots from the gel were subjected to protein identification using a nano-liquid chromatography system coupled to an ion-trap mass spectrometer equipped with an electrospray ionization nano-sprayer. We found that 10 protein spots were overexpressed in the serum samples from infected dogs compared with healthy dogs, which corresponded to three proteins: serotransferrin, serotransferrin isoforms X1, and hemopexin. Furthermore, analysis of the protein-protein interaction network confirmed that they strongly interacted with each other. This study suggests that high levels of serotransferrin and hemopexin are related to B. canis infection, making these proteins potential candidates for the development of diagnostic molecules or vaccines. Keywords: Babesia canis, blood parasite, diagnosis, proteomics.

The association between bacterial DNA in stifle joints, including those with cranial cruciate ligament rupture (CCLR) and medial patellar luxation (MPL), and osteoarthritis in dogs remains elusive. This study investigated the potential association between the detection of bacterial DNA and osteoarthritis in dogs using a broad-range polymerase chain reaction technique targeting the 16S ribosomal RNA gene. Synovial fluid (35 samples) and knee tissue samples (32 samples) were obtained from 35 dogs diagnosed with CCLR (n = 20; 11 males and nine females) or MPL (n = 15; five males and 10 females) who underwent a surgical operation between October 2014 and April 2015. Dogs with CCLR had a higher average osteoarthritis score than those with MPL (2.0 ± 0.9 vs. 0.5 ± 0.9; p = 0.005). Bacterial DNA was detected in the stifle joints of 60.71% of dogs with MPL. Pelomonas spp. (25.00%), Halomonas spp. (17.86%), and 5 other species (17.86%) were the most frequently identified bacteria. Bacterial DNA was detected in 41.03% of dogs with CCLR. Pelomonas spp. (15.38%), Sphingomonas spp. (10.26%), Halomonas spp. (5.13%), and 4 other species (10.26%) were the most frequently identified bacteria. No significant difference was observed in the prevalence of bacterial DNA obtained from tissue samples (46.88%) or joint fluid samples (51.43%). The presence of bacterial DNA was not associated with the type of knee injury (MPL or CCLR; p = 1.000). There was a higher prevalence of bacterial DNA in samples from dogs with moderate-to-severe osteoarthritis (94.44%) than in those with minimal osteoarthritis (41.18%), and a significant association between the presence of bacterial DNA and moderate-to-severe osteoarthritis was identified (p < 0.01). Dogs with moderate-to-severe osteoarthritis were more likely to have bacterial DNA in their stifle joints than those with no or minimal osteoarthritis. These findings provide valuable insight into the potential role of bacterial DNA in joint tissue or joint fluid and the development of osteoarthritis in dogs. Keywords: bacterial DNA, cranial cruciate ligament, osteoarthritis, stifle.

The dietary probiotics in sows during gestation to lactation period have gained considerable attention with respect to their beneficial effects on sows and their piglets’ performance and health. This study aimed to evaluate the effects of using probiotics in late-pregnant and lactating sows on milk quality, quantity, fecal microflora of sows, and growth performance of their offspring until weaning. Thirty-four sows were equally divided into two groups (control and treatment). Only those in the treatment group were fed 5 g of probiotics at 12 weeks of pregnancy, once daily for 7 weeks, until their piglets were weaned. Colostrum samples were collected at 3, 6, 12, and 24 h after farrowing and measured for immunoglobulin concentration. Percentages of fat, protein, and lactose in colostrum, colostrum production, total intake of immunoglobulin A (IgA), immunoglobulin G (IgG), fat, protein, and lactose, the change of fecal microflora of sows, and average daily gain of piglets were measured. The results showed that there were no significant differences in the concentrations of IgA, IgG, and IgM in colostrum and the percentages of fat, protein, lactose, solid-not-fat, and total solid in colostrum between the groups; however, the colostrum production at 24 h in the treatment group (6,075.29 mL) was higher than in the control group (4,809.54 mL). Higher total intakes of IgA and IgG as well as total intake of fat, protein, and lactose, particularly at 3 h after farrowing, were found in the treatment group. Probiotic supplementation remarkably altered the microbiota community at the phylum level. We found that Firmicutes and Bacteroidetes are the dominant phyla, present in the gut of more than 90% of pregnant and lactating sows. Changes in microbial proportions were observed due to the changes of pig production stage. The weaning weight of the treatment group was higher than in the control group (6.34 ± 1.71 vs. 4.84 ± 1.29 kg, respectively). Feeding of multi-species probiotic BACTOSAC-P™ during late pregnancy and lactation in sows positively influenced colostrum production. In this experiment, the use of BACTOSAC-P™ improved the yield of colostrum production. The high immunoglobulin concentration and high yield of the colostrum of sows with a diet supplemented with BACTOSAC-P™ significantly reduced piglet mortality during the suckling period. Furthermore, the probiotic diet induced changes in the fecal microbial population in sows by increasing the number of microorganisms from the Firmicutes phylum, which had positive effects on sow health and their piglets, leading to better piglet growth performance. Keywords: fecal microflora, lactation, probiotics, swine, weaning.

Enterotoxigenic Escherichia coli (ETEC) poses a substantial risk of neonatal diarrhea and post-weaning diarrhea among piglets, with F4+ ETEC strains emerging as a particularly challenging issue within the pig farming industry. This study aimed to introduce a straightforward approach for generating a crude extract of F4 fimbriae that shows promise as an antigenic determinant for potential vaccination strategies. A crude F4 fimbriae extract was obtained from F4+ ETEC using a combination of heat shock and homogenization techniques. Subsequently, three 4-week-old piglets were immunized with a primary dose of 150 μg and a booster dose 2 weeks later. Blood samples were collected to evaluate the level of serum F4-specific antibodies using an enzyme-linked immunosorbent assay. Analysis using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and liquid chromatography tandem-mass spectrometry techniques unveiled crucial insights into the composition of the crude F4 fimbriae extract. Notably, a distinct prominent band (~24 kDa) was identified, corresponding to the size of FaeG, the major subunit of F4 fimbriae. Regarding antibody response, there was a remarkable disparity between the levels of serum immunoglobulin (Ig)G and IgA antibodies targeting F4 compared with other E. coli strains (F18+ ETEC, F41+ ETEC, and F4−F18−F41− EC), as well as with the unvaccinated control group (p < 0.01). Specifically, the levels of IgG antibodies against other E. coli strains were also significantly higher than those observed in the unvaccinated control group (p < 0.05). Our findings suggest that the crude F4 fimbriae extracts obtained using our simple extraction method induce specific immune responses against F4+ E. coli and stimulate cross-immunity against other E. coli strains. Therefore, our method shows potential for use in future vaccine development against diarrhea in pigs caused by E. coli. Keywords: diarrhea, Escherichia coli, F4 fimbriae, piglets, vaccine.

Lumpy skin disease (LSD) is an emerging epidemic in livestock in Indonesia. It was first reported in the Indragiri Hulu Regency of Riau Province, which has more cases than the surrounding regencies. This study aimed to identify the risk factors and generate a logistic regression model and vulnerability map of LSD in the Indragiri Hulu Regency. We used a structured questionnaire to interview the case and control farm owners to evaluate the risk factors. We evaluated 244 samples, consisting of 122 case and control farm samples each. At the cattle farm level, the risk factor data related to LSD were analyzed using descriptive statistics, bivariate analysis with Chi-square, and odds ratio, while the logistic model was derived using multivariate logistic regression analysis. Using variables, such as the number of cases and risk factor variables included in the model logistic, and the temperature, humidity, and rainfall data from the Meteorology, Climatology, and Geophysical Agency, we analyzed the vulnerability map of LSD in the regency using scoring, weighting, and overlay methods. Ten significant risk factors were associated with LSD occurrence. The LSD model obtained from the logistic regression analysis was LSD (Y) = −3.92095 + 1.13107 (number of cattle >3) + 1.50070 (grazing cattle together with other farmers’ cattle) + 1.03500 (poor management of farm waste/dirt) + 2.49242 (presence of livestock collectors/traders near the farm location) + 1.40543 (introduction of new livestock) + 2.15196 (lack of vector control measures on the farm). The LSD vulnerability map indicated that the villages with high vulnerability levels were Rantau Bakung, Kuantan Babu, and Sungai Lala in the Rengat Barat, Rengat, and Sungai Lala subdistricts, respectively. We found 10 significant risk factors associated with LSD occurrence. The LSD model included the number of cattle (>3), cograzing with other farmers’ cattle, poor management of farm waste/dirt, the presence of livestock collectors/ traders near the farm, introduction of new livestock, and lack of vector control measures on the farm. The LSD vulnerability map indicated that villages with high vulnerability levels included Rantau Bakung in the Rengat Barat subdistrict, Kuantan Babu in the Rengat subdistrict, and Sungai Lala in the Sungai Lala subdistrict. Keywords: Indragiri hulu, lumpy skin disease, risk factors, vulnerability map.

The diagnosis of bovine brucellosis in animals vaccinated with strain-19 (S19) and Rose Bengal (RB)-51 strain vaccines can be misinterpreted due to false positives. This study aimed to compare diagnostic tests for detecting bovine brucellosis in animals vaccinated with S19 and RB51 vaccine strains. Two groups of 12 crossbred Holstein calves between 6 and 8 months of age were used. On day 0, blood samples were collected from the animals, and the competitive enzyme-linked immunosorbent assay was used for serological diagnosis of bovine Brucellosis. All animals tested negative. After the first blood collection, the animals were subcutaneously vaccinated: one group received the S19 vaccine and the other received the RB51 vaccine. From the 3rd month after vaccination, all animals were sampled. Sampling was repeated every 2 months until the 7th month. Serological diagnosis of bovine brucellosis was performed using RB, tube serum agglutination test (SAT), SAT with 2-mercaptoethanol (SAT-2Me), and fluorescence polarization assay (FPA). Animals vaccinated with S19 showed positive results with the RB, SAT, and SAT-2Me tests in all months of post-vaccination diagnosis. In animals vaccinated with S19, FPA showed positive results at months 3 and 5 and negative results at month 7, indicating that this test discriminates vaccinated animals from infected animals 7 months after vaccination. Rose Bengal, SAT, SAT-2Me, and FPA tests showed negative results in animals vaccinated with RB51 in all months of diagnosis. Animals vaccinated with S19 may test positive for brucellosis using RB, SAT, or SAT-2Me tests 7 months later. Fluorescence polarization assay is an optimal alternative for diagnosing animals in the field, thereby preventing false positives, and consequently, unnecessary confiscations of animals. Animals vaccinated with RB51 tested negative with RB, SAT, SAT-2Me, and FPA tests in all months of diagnosis, confirming that the tests are ineffective for diagnosing brucellosis caused by rough strains. Keywords: agglutination, bovine, brucellosis, vaccination.

Newcastle disease (ND) is a major viral disease of poultry worldwide. However, data on the molecular characterization of Newcastle disease virus (NDV) in Vietnam are limited. This study aimed to identify the molecular characteristics of NDV strains from the vaccinated chickens farmed in Northern Vietnam. We used reverse-transcription polymerase chain reaction (PCR), sequencing and phylogenetic analysis to characterize NDV strains from vaccinated chicken farms in Northern Vietnam. Seven out of 72 (9.7%) chicken tissue samples collected from seven chicken farms in the four cities/provinces in northern Vietnam were positive for the NDV genome by PCR method. The complete sequences of the fusion (F) and hemagglutinin-neuraminidase (HN) genes of NDVs isolated in the North of Vietnam from 2021 to 2022 were further evaluated. The results indicated that all seven Vietnamese isolates obtained were reported as virulent NDV strains with the amino acid (AA) sequence of the F0 protein proteolytic cleavage site motif (112RRRKRF117). Phylogenetic analysis revealed that they were grouped with other NDV class II from subgenotype VII.2, including the two previous Vietnamese NDV (2015), the Chinese (2017), and Southern African (2013) NDV strains. In addition, some AA substitutions were observed in the neutralizing epitopes of the F and HN proteins of the current Vietnamese NDV strains. The present findings provide useful information for future studies of the evolution of NDVs and improve strategies for ND-controlling programs in Vietnam. Keywords: genetic characterization, Newcastle disease virus, Vietnam, genotype VII.2, virulent strain.

Salmonella spp. is frequently found in the digestive tract of birds and reptiles and transmitted to humans through food. Salmonellosis is a public health problem because of pathogenicity variability in strains for virulence factors. This study aimed to identify the virulence genes in Salmonella isolates from humans, crocodiles, broiler cloacas, and broiler carcasses from two departments of Colombia. This study was conducted on 31 Salmonella enterica strains from humans with gastroenteritis (seven), crocodiles (seven), broiler cloacas (six), and broiler carcasses (12) from Tolima and Santander departments of Colombia, belonging to 21 serotypes. All samples were tested for Salmonella spp. using culture method on selective and non-selective mediums. Extraction of genomic DNA was performed from fresh colonies, DNA quality was verified by spectrophotometry and confirmed by amplification of InvA gene using conventional polymerase chain reaction (PCR). bapA, fimA, icmF, IroB, marT, mgtC, nlpI, oafA, pagN, siiD, spvC, spvR, spvB, Stn, and vexA genes were amplified by PCR. The most prevalent gene was bapA (100%), followed by marT (96.77%), mgtC (93.55%), and fimA (83.87%). Likewise, IroB (70.97%), Stn (67.74%), spvR (61.29%), pagN (54.84%), icmF (54.8%), and SiiD (45.16%) were positive for more than 50% of the strains. Furthermore, none of the isolates tested positive for the vexA gene. Salmonella isolates presented 26 virulence profiles. This study reported 14 virulence genes in Salmonella spp. isolates from humans with gastroenteritis, crocodiles, and broiler cloacas and carcasses. The distribution of virulence genes differed among sources. This study could help in decision-making by health and sanitary authorities. Keywords: broilers cloaca, carcasses, crocodiles, gastroenteritis human, polymerase chain reaction, virulence genes.

Parasitic diseases, including trematode invasions, result in losses to livestock in Indonesia, hindering the government’s efforts to achieve meat self-sufficiency. This study aimed to estimate the prevalence of Amphistomes and Fasciola in large ruminants reared by smallholder farmers. Fecal samples from 199 buffalo and cattle were collected from the districts of East Lampung (Lampung Province) and Lebak (Banten Province). Fecal samples were examined for the presence of trematode eggs using a sedimentation technique. Parasite invasion rate was 48.2% (95% confidence interval [CI]: 41.3%-55.2%). Rate of invasion was 63.3% (95% CI: 52.7%-73.9%) in Lampung and 38.3% (95% CI: 29.6%-47.0%) in Lebak-Banten. The prevalence of multiple invasions of both Amphistomes and Fasciola was 20% in buffalo and local cattle, whereas invasion rate was 12.8% in crossbred cattle. Invasion rate of Amphistomes alone was 27.1%, and that of Fasciola was 4.5%. A higher invasion rate of Amphistomes (29.8%) occurred in crossbred animals. There were no significant differences between age groups for trematode invasion. The Chi-square test showed that the prevalence of trematode invasion in females was significantly higher than in males (51.5% and 30.0%, respectively). Amphistomes more commonly infected females than males (29.0% and 16.7%, respectively). All breeds were vulnerable to invasion by both trematode species and single invasions with different invasion rates. These findings contribute to determining the magnitude of the disease and provide a basis for studies on prevention and treatment of trematode invasion. Keywords: amphistomes, Fasciola, Indonesia, large ruminants, prevalence.

Leptospirosis is considered a neglected tropical zoonosis in low-income countries due to surveillance system limitations and non-specificity of symptoms. Humans become infected through direct contact with carrier animals or indirectly through Leptospira-contaminated environments. Conventionally, equines have been considered an uncommon source of leptospirosis, but recent publications in Latin America suggest that their role in the maintenance and dispersion of the bacteria could be more relevant than expected, as horses are susceptible to a wide variety of zoonotic Leptospira spp. from domestic and wild animals with which they share the environment. A systematic review of the published literature was conducted to compile the available information on Leptospira spp. in Ecuador, with a special focus on equine leptospirosis, to better understand the epidemiology of the bacterium and identify possible knowledge gaps. A systematic review of the published literature was conducted in PubMed, SciELO and Web of Science databases to compile the available information on Leptospira spp. in Ecuador, with a special focus on equine leptospirosis, to better understand the epidemiology of the bacterium. We used a combination of the terms (Leptospira OR Leptospirosis) AND Ecuador, without restrictions on language or publication date. Our literature review reveals that published scientific information is very scarce. Eighteen full-text original scientific articles related to Leptospira or leptospirosis cases in Ecuador were included in the systematic review. Most of the studies reported data obtained from one of the four regions (Coast), and specifically from only one of the 24 Provinces of Ecuador (Manabí), which evidence a large information bias at the geographical level. Furthermore, only the studies focused on humans included clinical signs of leptospirosis and there is only one study that analyzes the presence of Leptospira spp. in water or soil as a risk factor for pathogen transmission. Finally, only one study investigated Leptospira in horses. Since sentinel species can provide useful data on infectious diseases when epidemiologic al information is lacking, and horses could be considered excellent sentinel species to reveal circulating serovars, we propose developing a nationwide surveillance system using horses. This cost-effective epidemiological survey method provides a baseline for implementing specific prevention and control programs in Ecuador and neighboring developing countries. Keywords: Ecuador, equine leptospirosis, neglected tropical zoonosis, one-health, sentinel species, systematic review.

Bovine tuberculosis (bTB) is an infectious disease of cattle, mainly caused by Mycobacterium bovis. This study aimed to evaluate the efficacy of the interferon-gamma (IFN-γ) assay and single-intradermal comparative tuberculin test (SICTT) in detecting bTB. In an earlier study, 150 positive, 83 inconclusive, and 480 negative animals from 24 cattle herds were screened using SICTT. From these groups, 125 positive, 17 inconclusive, and six negative animals were subsequently verified using the IFN-γ assay. Single-intradermal comparative tuberculin test outcomes were interpreted according to standard guidelines, whereas blood samples were collected and stimulated with purified protein derivatives. Sandwich enzyme-linked immunosorbent assay was used to measure secreted IFN-γ. Concordant and Bayesian latent class analyses were performed to evaluate test performance. Results from the IFN-γ assay revealed that 83.2%, 64.7%, and 16.67% of the animals were positive in the SICTT-positive, inconclusive, and negative animal categories, respectively. Sensitivity (SE) and specificity (SP) of SICTT were 83.9% (95% confidence interval [CI]: 77.4-90.1) and 95.7% (95% CI: 86.9-99.7), respectively. Sensitivity and SP for the IFN-γ assay were 78.9% (95% CI: 71.9-85.4) and 83.9% (65.9-95.9), respectively. The use of both tests in parallel increases the SE of bTB detection (~94%), compared with SICTT alone. Use of the IFN-γ assay with SICTT in parallel, predominantly on cattle demonstrating an inconclusive SICTT outcome, boosts bTB detection rate in low resource settings. Keywords: Bayesian latent class analysis, farmed dairy cattle, gamma-interferon assay, single-intradermal comparative tuberculin test.

Stomoxys and tabanid flies are of medical and veterinary importance because they play crucial roles in disease transmission as mechanical vectors of various hemopathogens. However, its role as a hemoprotozoan parasite vector in horse farms has not been studied. Therefore, we investigated the occurrence of hemoprotozoan parasites belonging to the genera Babesia, Theileria, and Trypanosoma in Stomoxys and tabanid flies using conventional polymerase chain reaction (PCR) and DNA sequencing. All samples (Stomoxys and tabanid flies) were collected using an Nzi trap for three consecutive days each month from November 2022 to March 2023. The flies were morphologically identified to the species level and separated according to sex. Individual (for tabanid flies) or pooled samples (consisting of three specimens of Stomoxys flies of the same species and sex collected from the same site) were used for DNA extraction. Conventional PCR was used to screen for hemoprotozoan parasite DNA, followed by Sanger sequencing to identify the species. In total, 189 biting flies were collected, including four species of Stomoxys (Stomoxys bengalensis, Stomoxys calcitrans, Stomoxys indicus, and Stomoxys sitiens) and five species of tabanids (Atylotus cryptotaxis, Chrysops dispar, Tabanus megalops, Tabanus mesogaeus, and Tabanus rubidus). Stomoxys calcitrans was the most prevalent species, accounting for 58.7% (n = 111) of the collected flies. Ten (12.4%) of the 81 samples (individuals and pools) analyzed by PCR were positive for the 18S rRNA gene of the Theileria/Babesia species. Trypanosoma DNA was not detected in any sample. After performing Basic Local Alignment Search Tool searches and a phylogenetic analysis, only six samples (7.4%), including S. calcitrans (n = 2), S. sitiens (n = 2), T. megalops (n = 1), and A. cryptotaxis (n = 1), were found to be infected with Theileria sinensis. Furthermore, apicomplexan parasites, namely, Mattesia spp. and Colpodella spp., were found on S. indicus, the fungus Meira spp. was found on S. calcitrans, and the pathogenic green alga Helicosporidium spp. was found on A. cryptotaxis. This study is the first to report a variety of Stomoxys and tabanid flies collected from horse farms in Thailand, which were found to be infected with Theileria and Colpodella species that affect mammals, suggesting that Stomoxys and tabanid flies can be used to confirm the presence of hemoprotozoan parasites in the study area. Understanding the presence of hemoprotozoa in flies could help design vector control programs and manage various diseases in the study area. Keywords: hemoprotozoan parasites, Stomoxys flies, tabanid flies, Thailand, vector.

Staphylococci, including Staphylococcus aureus, Staphylococcus chromogenes, and Staphylococcus haemolyticus, are significant bacteria that induce bovine mastitis, primarily because they can form biofilms in bovine teat canals. This study aimed to investigate the efficacy of Piper betle extract and a bovine teat dipping solution containing P. betle extract (BSP) against these mastitis-causing staphylococci. BSP was prepared using P. betle extract as the bioactive compound. The antibacterial activity of the plant extract and BSP against the pathogens was investigated using a broth microdilution method. The activity of the extract and BSP against the pathogen biofilms was also determined. A stability test was performed to observe the pH, color, turbidity, homogeneity, precipitation, and separation of BSP stored at 4°C and 25°C for up to 4 weeks. The extract exhibited potent antibacterial activity against S. aureus and S. haemolyticus, with similar values for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) ranging from 0.03 mg/mL to 0.125 mg/mL. The MIC and MBC values of the extract against S. chromogenes were 0.5-1 mg/mL and 0.5-2 mg/mL, respectively. Moreover, BSP exhibited MIC and MBC values of 12.5–50 v/v against all tested staphylococci isolates. When used at 1/2 and 1/4 × MIC, the extract and BSP significantly inhibited the formation of staphylococcal biofilms (p < 0.05) in the tested strains. The results indicated that treatment with 1/2 × MIC of the extract and BSP resulted in biofilm inhibition ranging from 30%–66% and 19%–39%, respectively. Furthermore, the extract at 16 × MIC eliminated 54%-86% of established mature isolate biofilms, whereas BSP removed 41%-61% of mature biofilm viability. Storage of BSP at 4°C did not change the factors associated with stability from the 1st to 4th week. These findings suggest that BSP may exhibit potential medicinal benefits in inhibiting the growth and biofilm formation of mastitis-inducing staphylococci in bovines. Keywords: antibacterial activity, biofilms, bovine teat dipping solution, Piper betle extract, staphylococci.

In recent years, artificial intelligence (AI) has become increasingly necessary in the life sciences, particularly medicine and healthcare. This study aimed to systematically review the literature and critically analyze multiple databases on the use of AI in veterinary medicine to assess its challenges. We aim to foster an understanding of the effects that can be approached and applied for professional awareness. This study used multiple electronic databases with information on applied AI in veterinary medicine based on the current guidelines outlined in PRISMA and Cochrane for systematic review. The electronic databases PubMed, Embase, Google Scholar, Cochrane Library, and Elsevier were thoroughly screened through March 22, 2023. The study design was carefully chosen to emphasize evidence quality and population heterogeneity. A total of 385 of the 883 citations initially obtained were thoroughly reviewed. There were four main areas that AI addressed; the first was diagnostic issues, the second was education, animal production, and epidemiology, the third was animal health and welfare, pathology, and microbiology, and the last was all other categories. The quality assessment of the included studies found that they varied in their relative quality and risk of bias. However, AI aftereffect-linked algorithms have raised criticism of their generated conclusions. Quality assessment noted areas of AI outperformance, but there was criticism of its performance as well. It is recommended that the extent of AI in veterinary medicine should be increased, but it should not take over the profession. The concept of ambient clinical intelligence is adaptive, sensitive, and responsive to the digital environment and may be attractive to veterinary professionals as a means of lowering the fear of automating veterinary medicine. Future studies should focus on an AI model with flexible data input, which can be expanded by clinicians/users to maximize their interaction with good algorithms and reduce any errors generated by the process. Keywords: artificial intelligence, Cochrane study, criterion, extracted data, heterogeneity, systematic review.

Seasonal changes, especially temperature and photoperiod, are well-known determining factors of swine reproductive capacity, but the underlying mechanisms remain unknown. This study aimed to investigate the effect of age and seasonal variations on boar seminal plasma steroids (dehydroepiandrosterone [DHEA], cortisol [CORT], and testosterone [TEST]) over 1 year. Four commercial hybrid adult boars (Large White × Duroc), aged between 12 and 44 months, were repeatedly evaluated at the Department of Veterinary Medical Sciences of the University of Bologna. Daily temperature and light hours relating to the collection date were considered for each observation within the four astronomical seasons: Winter, spring, summer, and autumn. Hormones were quantified using radioimmunoassay. The association between seasonal factors and hormone concentrations was evaluated using linear regression models. Univariate models were estimated for each hormone to assess the influence of the independent variables; two multivariate models were assessed to evaluate the effect of temperature and daylight hours, including boar and season factors. Age significantly affected all analyzed hormones (CORT p < 0.0001; DHEA p < 0.0001; and TEST p < 0.0001). The highest average levels were found for each hormone during summertime, suggesting a positive correlation between steroid concentrations with temperature and light hours. The results of this study support the hypothesis that the increase in external temperature and light hours is somehow associated with higher levels of steroid concentrations in the seminal plasma of in-housed boars. These findings may help further investigate seasonal fluctuations in reproductive outcomes, which are well-known for porcine species. Keywords: boar, ejaculate, reproduction, seasonal parameters, steroids, swine.

Malaysia has more than 630 culturists who are involved in the ornamental fish industry and culture 250 species, including local and exotic species. Among these viruses, megalocytiviruses have been associated with severe systemic diseases and economic losses in ornamental fish. The intensity of Megalocytivirus infection in Pterophyllum scalare in Malaysia remains unknown. Thus, this study aimed to investigate the occurrence of Megalocytivirus while discovering its associated risk factors and the genotypes of its causative agents in an ornamental fish farm in Malaysia. Seven broodstock pairs of P. scalare were used in this study to follow the life stages of fish, from egg to market size. Water samples and other samples, such as mucus swabs, gill swabs, P. scalare eggs, fries, juveniles, snails, snail eggs, live feed (Tubifex worms and Moina spp.), sediment samples, and wild fish, were collected periodically for initial environmental sampling from day 0 to day 60. Nested polymerase chain reaction amplifications were performed for megalocytivirus-related sequences. The phylogenetic tree, including the sampled causative agents of megalocytiviruses, was inferred from the major capsid protein genes of all known Iridoviridae species. Pearson’s correlation coefficients were calculated to determine the strength of the correlation between the presence of megalocytiviruses in P. scalare samples and the associated risk factors. A total of 312 out of 935 pooled and individual samples tested positive for the presence of Megalocytivirus-related sequences, except snail eggs and wild fish (Poecilia reticulata). No clinical symptoms were observed in any fish samples. Megalocytivirus-associated viruses detected in water samples indicate horizontal transmission of the virus. All the nucleotide sequences found in this study had high nucleotide identities of 95%–99 % and were closely related to Megalocytivirus genotype I infectious spleen and kidney necrosis virus. Risk factors associated with Megalocytivirus include water temperature, dissolved oxygen (DO), pH, ammonia, nitrate, nitrite, and the life stages of P. scalare. High Megalocytivirus infection was detected when the water temperature, DO, and pH were high in P. scalare, high water temperature and nitrate in the water samples, and the same rate of Megalocytivirus infection in P. scalare fry and juveniles. This is the first study to confirm the existence of different possible routes of megalocytivirus distribution in ornamental fish farms in Malaysia. Nevertheless, the connection between the mode of transmission and the risk factors for this virus needs to be explored further to recognize the evolution and potential new host species. Keywords: epidemiology, genotype I, infectious spleen and kidney necrosis virus, Megalocytivirus, ornamental fish, risk factors.

The heart conduction system is responsible for the occurrence of various types of cardiac arrhythmia. This study aimed to histologically and morphometrically describe damaged cardiac nodes during acute myocardial infarction and to compare them with normal tissues in dogs and horses. This study describes the morphometry of cardiac nodes in five dogs and five elderly horses that succumbed to sudden cardiac death (SCD). A computerized morphometric study was conducted to determine the number of cells composing the nodes, different shape and size parameters of nodes, and their relationship with degenerative changes due to cardiac conditions. In both species, the sinoatrial node (SAN) was ovoid in shape whereas the atrioventricular node (AVN) was pyramidal in shape. The percentage of collagen fibers inside the SAN of dogs (47%) and horses (50%) was found to be higher than that of cells. In contrast, the percentage of cells in the AVN of dogs (24%) and horses (16%) was higher than that of connective tissues. In the SAN, the area (p = 0.09), maximum diameter (<0.001), and mean diameter (0.003) of P cells were larger in dogs than in horses. Overall, the SAN cells and surrounding cardiomyocytes in dogs and horses as well as the AVN cells in dogs that succumbed to SCD decreased in size compared with those in normal hearts. Keywords: atrioventricular node, dog, horse, myocardial infarction, sinoatrial node.