Cattle Feces Research Articles

Assessment of Bacterial Contamination and Antimicrobial Resistance of Escherichia coli Isolates from Slovak Dairy Farms

The conditions in livestock housing are suitable for the survival of airborne microorganisms, mainly due to high temperatures, humidity, and the presence of organic material. The total count of airborne bacteria concentrations in cattle farms ranged from 3.01 log10 CFU/mL to 6.90 log10 CFU/mL; for coliform bacteria, they were from 2.18 log10 CFU/mL to 3.34 log10 CFU/mL; and for molds, they ranged from 3.00 log10 CFU/mL to 4.57 log10 CFU/mL. Bacteria resistant to antimicrobial substances and resistance genes can be spread on animal farms. Antimicrobial resistance in ubiquitous Escherichia coli isolated from cattle feces was investigated. Minimum inhibitory concentration (MIC) testing was utilized to identify phenotypic resistance profiles, and the PCR method was employed to detect the presence of resistant genes. A higher percentage of resistance was found to amikacin (65%), tetracycline (61%), streptomycin (56%), ampicillin (55%), and nalidixic acid (45%). Multidrug resistance was determined in up to 64.3% of the isolates studied. The most widespread resistance genes were blaTEM (85.7%), sul2 (66.7%), tetB (52.38%), and sul1 (47.6%). We found that 4.8% of the E. coli isolates had the blaCMY gene. We found that, despite phenotypic resistance, E. coli isolates do not necessarily carry genes conferring resistance to that particular antimicrobial agent.

Biomethane Potential of Beef Cattle Slaughterhouse Waste and the Impact of Co-Digestion with Cattle Feces and Swine Slurry

Slaughterhouse waste (SW) poses significant environmental challenges due to its complex composition, but anaerobic digestion offers a way to recover valuable biogas from SW. This study investigated the anaerobic co-digestion of beef cattle slaughterhouse waste (BCSW) with either cattle feces (CF) or swine slurry (SS). The biomethane potential, maximum methane yield (Mmax), lag phase duration, and effective digestion time (Teff) for the individual substrates and the combinations were analyzed. BCSW alone exhibited Mmax of 578.5 Nml CH4/g VSadded with a lag phase of 11 days, while CF and SS alone exhibited Mmax of 397.2 and 289.8 Nml CH4/g VSadded, respectively. Co-digestion of BCSW and SS resulted in Mmax increase of 48–75.5%, with negligible effects on Teff compared to solitary SS digestion. Similarly, co-digestion of BCSW and CF increased Mmax by 6.2–40.4%, with no significant impact on Teff compared to solitary CF digestion. However, both co-digestions led to a reduction in Mmax (12.1–27%) when compared to BCSW digestion alone. Co-digestion with SS shortened the lag phase duration by 2.8–7.8 days and accelerated Teff by 5.8–8.3 days due to SS’s high concentrations of essential micronutrients like cobalt and nickel which aid digestion. This study concluded that co-digestion of BCSW with SS is an effective strategy for enhancing methane production and digestion efficiency, offering a viable approach for proper disposal of BCSW while improving biogas output.

Campylobacter californiensis sp. nov., isolated from cattle and feral swine.

Nine Campylobacter strains were isolated from cattle and feral swine faeces: three were recovered during a 2007 Campylobacter-associated outbreak linked to a dairy, and the other six were isolated during a 2009-2010 survey of farms and ranches in Central California. The species identification of these strains could not be determined by 16S rRNA gene sequencing but were most similar to Campylobacter concisus and Campylobacter mucosalis. Additional atpA typing indicated that the nine strains composed a discrete novel clade related to C. concisus and C. mucosalis. A polyphasic study was undertaken here to clarify their taxonomic position. Phylogenetic analyses were performed based on 16S rRNA gene sequences and the concatenated sequences of 330 core genes. The core gene analysis placed the nine strains into a clade well separated from the other Campylobacter taxa, indicating that these strains represent a novel Campylobacter species. Pairwise digital DNA-DNA hybridization and average nucleotide identity values between these strains and other campylobacters are lower than 16 and 73%, respectively, further supporting their placement into a novel taxon. Standard phenotypic testing was performed. All strains are microaerobic or anaerobic, motile, Gram-negative, slightly-curved rods that are oxidase positive but catalase negative. Strains can be distinguished from the other catalase-negative Campylobacter species using phenotypic markers such as motility, oxidase activity, cephalothin resistance, hippuricase activity, growth at 30 °C, and α-haemolysis. The data presented here show that these strains represent a novel species within Campylobacter, for which the name Campylobacter californiensis sp. nov. (type strain RM6914T=LMG 32304T=CCUG 75329T) is proposed.

Environmental contamination of arsenic: pathway analysis through water-soil-feed-livestock in Nadia District (India) and potential human health risk.

This study investigated arsenic (As) concentrations in diverse environmental components and their potential impact on the health risks faced by residents of the arsenic (As)-contaminated Nadia district in West Bengal, India. A random selection of 182 cattle and 255 goats from 40 livestock farmers in the district revealed that both animals and humans were naturally exposed to elevated arsenic levels through contaminated drinking water, foods, grasses, concentrate feeds, various fodder tree leaves, and other food/feed resources. The mean As concentration in roughages (483.18µg/kg DM) was significantly higher (p < 0.001) than in tree leaves (391.53µg/kg DM), and concentrate feed/ingredients (186.66µg/kg DM). Pond water exhibited higher arsenic levels (106.11µg/L) compared to shallow tube well water (47.96µg/L) and deep tube well water/tap water (10.64µg/L and 10.04µg/L, respectively). The mean arsenic concentration in soils DM of fodder fields, crop fields, and grassland was 10.25, 10.58, and 10.20mg/kg, respectively. It was observed that protein-rich feeds had lower levels of arsenic accumulation (p < 0.048), while fiber-rich feeds containing more cellulose, hemicellulose, and lignin had higher arsenic levels (p < 0.017). Goats consumed 73.46% more arsenic per kg body weight compared to dairy cows. Although chronic and sub-chronic arsenic exposure in the district did not typically manifest symptoms or visible signs in ruminant animals, concentrations in the hair and feces of both cattle and goats exceeded normal values. Cattle feces had significantly higher arsenic (410.43µg/kg DM) levels (p < 0.001) than goat feces (227.00µg/kg DM), and arsenic concentration in cattle hair (1917.74µg/kg DM) was also significantly greater (p < 0.001) than goat hair (1435.74µg/kg DM). Arsenic levels in milk samples from both species were below 10µg/kg. Liver (356.02µg/kg DM) and kidney (317.22µg/kg DM) contained significantly higher (p < 0.001) levels of arsenic compared to muscle (204.23µg/kg DM), and bone (161.98µg/kg DM) in local meat-type adult male goats. The skin accumulated the highest amount of arsenic (576.24µg/kg DM) among the non-edible parts of the goat carcass. The cumulative cancer risk value for adults was 4.96 × 10-3, exceeding the threshold value (1 × 10-6). This suggests a significant risk of cancer development for the population in arsenic-affected areas. Non-cancer risks (hazard indexes) were estimated at 11.01 for adults. Our observations revealed that the highest bioaccumulation of arsenic occurred in the hair of cows, and goats in the examined localities. The biotransformation factor (BTF) for hair was much higher compared to other excreted samples from both species. The calculated BTF followed the order: hair > feces > milk for cows and goats. Livestock farmers in Nadia district are advised to carefully select feed resources, prioritizing those high in crude protein and low in neutral detergent fiber, and they should provide drinking water from deep aquifers to ensure the safety of milk and meat for human consumption.

192 Prediction of fecal starch content of fattening cattle using near-infrared spectroscopy and machine learning

Abstract Starch content in cattle feces can be used to predict starch digestibility. Near-infrared spectroscopy (NIRS) can be used to rapidly and non-chemically predict fecal starch content after a calibration model has been consequentially developed. Recent advancements in machine learning have introduced potent methods for developing optimized models. In addition, diverse preprocessing methods, such as scatter correction and smoothing, are available to eliminate irrelevant variance sources. This study aimed to identify optimal spectral preprocessing and regression methods for predicting fecal starch content using NIR spectra. NIR spectra were measured for 196 fecal samples collected from 9 fattening cattle farms. Fecal starch content on a DM basis was measured using enzymatic reactions and colorimetry. The 196 samples were divided into a calibration set (n = 123), a validation set (n = 29) comprising four farms, and an external validation set (n = 44) comprising five farms that differed from the calibration set. Based on the calibration set, two preprocessing methods were selected using the partial least squares (PLS) model. In method 1, the calibration models were developed by a grid search 5-fold cross validation on the 1,152 preprocessed calibration sets, and preprocessing with the lowest root mean square error (RMSE) of cross-validation among these calibration sets was selected. In method 2, the original calibration set (n = 123) was divided into a calibration set (n = 116) and a preprocessing selection set (n = 7) with different neutral detergent fiber and similar starch contents. After preprocessing, calibration models were developed as described in method 1. The preprocessing selection sets were then predicted by the developed models, and preprocessing with the lowest RMSE among these preprocessing selection sets was selected. Using the preprocessing methods selected in methods 1 and 2, the validation and external validation sets were predicted using seven regression models. As a result, the preprocessing selected by method 1 was in the wavelength range of 400–2500 nm, with robust normal variate (RNV) scatter correction and first-order derivative Savitzky-Golay filtering (2nd order polynomial, 2-nm window size). The preprocessing selected by method 2 had a wavelength range of 1500–2500 nm, RNV scatter correction, and 2nd-order derivative Savitzky-Golay filtering (3rd order polynomial, 25-nm window size). Among the seven regression models, the Lasso model was the most accurate for prediction. The Lasso models based on the preprocessing methods selected by methods 1 and 2 had RMSE of external validation = 2.054% of DM and 1.540% of DM, respectively. The feature importance of the Lasso model based on preprocessing method selected by methods 2 was higher at wavelengths of 1775 nm and 2325 nm. In conclusion, seven models were developed and evaluated based on diverse preprocessing methods using NIRS. The Lasso model exhibited the highest accuracy.

Phenotypic and genomic comparison of three human outbreak and one cattle-associated Shiga toxin-producing Escherichia coli O157:H7.

Foodborne illness has major impacts on global health and imposes financial hardships on food industries. Escherichia coli serotype O157:H7 is associated with foodborne illness. Cattle feces are a source of E. coli O157:H7, and routine surveillance has led to an abundance of E. coli O157:H7 genomic data. The relationship between E. coli O157:H7 genome and phenotype is not clearly discerned for cattle colonization/shedding and improved understanding could lead to additional strategies to limit E. coli O157:H7 in the food chain. The goal of the research was to evaluate genomic and phenotypic attributes of E. coli O157:H7 associated with cattle colonization and shedding, environmental persistence, and human illness. Our results indicate variations in biofilm formation and in vitro cellular adherence was not associated with differences in cattle colonization or shedding. Overall, processes involved in cattle colonization and various phenotypes in relation to genotype are complex and remain not well understood.

Proposal of Lactobacillus amylovorus subsp. animalis subsp. nov. and an emended description of Lactobacillus amylovorus.

Seven novel lactic acid bacterial strains (BF125T, BF186, TKL145, YK3, YK6, YK10 and NSK) were isolated from the fresh faeces of Japanese black beef cattle and weanling piglets, spent mushroom substrates, or steeping water of a corn starch production plant. These strains are rod-shaped, Gram-stain-positive, non-motile, non-spore-forming, catalase-negative, cytochrome oxidase-negative, facultatively anaerobic, and homofermentative. Strain BF125T did not produce any gas from glucose; both d- and l-lactate were produced as end-products of glucose (D/L, 40 : 60). Growth occurred at 30-45 °C (optimum, 37 °C), pH 5.0-8.0 (optimum, pH 6.0), and with NaCl concentration of 1.0-3.0% (w/v). The G+C content of genomic DNA of strain BF125T was 37.8 mol% (whole-genome analysis). The major fatty acids were C16 : 0, C18 : 1 ω9c, C19 cyclopropane 9, 10, and summed feature 10. The 16S rRNA gene in strain BF125T showed high similarity to that of the type strain of Lactobacillus amylovorus (99.93%), and the other isolates were also identified as L. amylovorus based on these similarities. A phylogenetic tree based on the core genomes of L. amylovorus strains (n=54), including the seven isolates, showed that they could be divided into two clusters. Strains YK3, YK6, YK10, and NSK were in the first cluster, along with the type strain DSM 20531T, while the second cluster included isolates BF125T, BF186, TKL145, and other strains isolated from various animal origins. Phenotypic differences in fermentability were observed for lactose, salicin, and gentiobiose between these two groups. The intergroup digital DNA-DNA hybridization values (72.9-78.6%) and intergroup average nucleotide identity values (95.64-96.92%) were comparable to values calculated using datasets of other valid subspecies of the genus (ex-) Lactobacillus. In light of the physiological, genotypic, and phylogenetic evidence, we propose a novel subspecies of L. amylovorus, named Lactobacillus amylovorus subsp. animalis subsp. nov. (type strain BF125T=MAFF 212522T=DSM 115528T). Our findings also led to the automatic creation of Lactobacillus amylovorus subsp. amylovorus subsp. nov. and an emended description of the species L. amylovorus.

Occurrence of Cryptosporidium Oocysts in Faeces, Milk and Water Sources in Sedentary Fulani Herds in Selected Local Government Areas of Kaduna State, Nigeria

Abstract Cryptosporidiosis is a neglected tropical zoonotic disease, commonly associated with a diarrhoea, caused by a protozoan parasite of the genus Cryptosporidium. The aim of this study was to determine the occurrence of Cryptosporidium oocysts in faeces, milk, and water sources, and its associated risk factors in sedentary Fulani herds. Seven hundred and fifty faecal samples (240, 180, 240 and 90 from cattle, sheep, goats and dogs, respectively), and 120 samples each, of milk and water were collected for this study. Faecal samples were concentrated by formol ether concentration technique, then stained using Modified Ziehl Neelsen staining method, to detect Cryptosporidium oocysts. Cryptosporidium oocysts in milk and water samples were also concentrated and stained with same stain. The prevalence of Cryptosporidium oocysts in (cattle, dog, sheep and goat faeces were 18.3 %, 15.6 %, 13.9 % and 4.2 %, respectively.), Significantly higher prevalence was observed in calves ≤ 1 year (P = 0.002), female cattle (P = 0.007) and animals with poor body condition (P &lt; 0.001) than in the other groups. Goats with poor body condition scores had significantly higher prevalence (P = 0.008) than other groups while dogs ≤ 6 months of age also had significantly higher prevalence (P = 0.03) than older dogs. The prevalence of Cryptosporidium oocysts in bulk cow milk was 11.7 % while a prevalence of 13.3 % was observed in the water bodies sampled. Hence the findings of this study are of great public health significance, therefore, inhabitants of these LGAs should be informed and educated on the need for improvement of sanitary measures during the course of milking these animals, and the need for adequate pasteurization, of milk before consumption.

Evidence for the occurrence of Acinetobacter faecalis in cattle feces and its emended description

This study provides an emended description of Acinetobacter faecalis, a species previously described based on a single isolate (YIM 103518T) from elephant feces in China. Our emended description is based on 15 novel isolates conspecific with the A. faecalis type strain, obtained from eight cattle farms in the Czech Republic. The A. faecalis strains have relatively small genomes (≈2.5−2.7 Mbp), with a GC content of 36.3−36.7 mol%. Core genome-based phylogenetic analysis showed that the 15 strains, together with the type strain of A. faecalis, form a distinct and internally coherent phylogroup within the genus. Pairwise genomic ANIb values for the 16 A. faecalis strains were 97.32−99.04 %, while ANIb values between the genomes of the 16 strains and those of the other Acinetobacter spp. were ≤ 86.2 %. Analysis of whole-cell MALDI-TOF mass spectra supported the distinctness and cohesiveness of the taxon. The A. faecalis strains could be differentiated from the other validly named Acinetobacter spp. by the absence of hemolytic activity along with their ability to grow at 37 °C and on L-aspartate, ethanol, and L-glutamate but not at 41 °C or on adipate or 2,3-butanediol. Reduced susceptibility to sulfamethoxazole, trimethoprim and/or streptomycin was shown in eight strains, along with the presence of corresponding antibiotic resistance genes. In conclusion, this study provides a comprehensive description of A. faecalis and demonstrates its occurrence in cattle feces. Though the ecological role of A. faecalis remains unknown, our results show its ability to acquire antibiotic resistance genes, likely as an adaptation to antibiotic selection pressure in livestock farms.

Metagenomic insights into resistome, mobilome and virulome in different fecal waste

Fecal waste is a significant source of antimicrobial resistance (AMR) pollution and provides valuable insights into the AMR development in animal and human populations within the “One health” framework. Various genetic elements, including antibiotic resistance genes (ARGs), biocide and metal resistance genes (BMGs), mobile genetic elements (MGEs), and virulence factor genes (VFGs), are crucial AMR risk determinants (ARDs). However, few studies focused on compositional characteristics of ARDs in different feces. Here, we analyzed 753 public metagenomes from human, pig, chicken, and cattle feces, revealing significant differences in ARD richness and abundance across fecal types, notably lowest in cattle samples. Tetracycline, multi-metal, and -biocide resistance genes were dominant resistome. A few core genes contributed to 25.6%–91.1% of gene abundance, and their correlations were stronger in cattle samples. Procrustes analysis showed that microbial composition had higher correlations with ARGs (M2 = 0.579) and BMGs (M2 = 0.519). Gammaproteobacteria was identified as major ARD-hosts especially in human and pig feces, and they mainly carried multi-resistance genes. MGEs exhibited direct positive effects on ARGs and BMGs, indirectly impacting VFGs. Utilizing random forest methods, we identified 42 indicator genes for tracking AMR pollution originating from fecal sources in the environments. This study offers new insights into understanding and controlling the AMR pollution of fecal waste from human and food animals.

Prevalence and antimicrobial susceptibility profile of Salmonella isolated from vegetable farms fertilized with animal manure in Addis Ababa Ethiopia

The resistance of foodborne pathogens to antimicrobial agents is a potential danger to human health. Hence, establishing the status of good agricultural practices (GAPs) and the antimicrobial susceptibility of major foodborne pathogens has a significant programmatic implication in planning interventions. The objective of this study was to assess the gap in attaining GAP and estimate the prevalence and antimicrobial susceptibility profile of Salmonella in vegetable farms fertilized with animal manure in Addis Ababa, Ethiopia. A total of 81 vegetable farms from four sub-cities in Addis Ababa were visited, and 1119 samples were collected: soil (n = 271), manure (n = 375), vegetables (n = 398), and dairy cattle feces (n = 75). Additional data were collected using a structured questionnaire. Isolation of Salmonella was done using standard microbiology techniques and antimicrobial susceptibility testing was conducted using disk diffusion assays. Carriage for antimicrobial resistance genes was tested using polymerase chain reaction (PCR). Among the 81 vegetable farms visited, 24.7% used animal manure without any treatment, 27.2% used properly stored animal manure and 80.2% were easily accessible to animals. The prevalence of Salmonella was 2.3% at the sample level, 17.3% at the vegetable farm level, and 2.5% in vegetables. The highest rate of resistance was recorded for streptomycin, 80.7% (21 of 26), followed by kanamycin, 65.4% (17 of 26), and gentamicin, 61.5% (16 of 26). Multidrug resistance was detected in 61.5% of the Salmonella isolates. Vegetable farms have a gap in attaining GAPs, which could contribute to increased contamination and the transfer of antimicrobial resistance to the vegetables. The application of GAPs, including proper preparation of compost and the appropriate use of antimicrobials in veterinary practices, are recommended to reduce the emergence and spread of antimicrobial resistance.

Isolation of microorganisms from the feces of Kitasato Yakumo beef cattle as bioactive natural product producers.

We envisioned that the rumen of Kitasato Yakumo beef cattle would contain unique microorganisms which produce bioactive compounds as their defense response to the external environment. The variety of microorganisms were collected from the feces of Kitasato Yakumo beef cattle. We evaluated the biological activity of the culture broth of the isolated strains, proving the utility of our approach.

A newly identified IncY plasmid from multi-drug-resistant Escherichia coli isolated from dairy cattle feces in Poland.

This study reveals the identification of new strains of antibiotic-resistant Escherichia coli in cattle manure from a dairy farm in Poland, offering critical insights into the spread of drug resistance. Through whole-genome sequencing, researchers discovered novel plasmids within these bacteria, which carry genes resistant to multiple antibiotics. These findings are particularly alarming, as these plasmids can transfer between different bacterial species, potentially escalating the spread of antibiotic resistance. This research underscores the vital connection between the health of humans, animals, and the environment, emphasizing the concept of One Health. It points to the critical need for global vigilance and strategies to curb the proliferation of antibiotic resistance. By showcasing the presence of these strains and their advanced resistance mechanisms, the study calls for enhanced surveillance and preventive actions in both agricultural practices and healthcare settings to address the imminent challenge of antibiotic-resistant bacteria.

The role of ruminant urine and faeces in the recycling of nutrients by forages

This study addresses the effect of using animal excreta on the nutritional content of forages, focusing on macro- and micro-element concentrations (nitrogen; N, phosphorus; P, sulphur; S, copper; Cu, zinc; Zn, manganese; Mn, selenium; Se) from animal feed to excreta, soil, and plants. Data were collected from pot and field trials using separate applications of sheep or cattle urine and faeces. Key findings indicate that soil organic carbon (SOC) and the type of excreta significantly influences nutrient uptake by forages, with varied responses among the seven elements defined above. Although urine contributes fewer micronutrients compared to faeces (as applied at a natural volume/mass basis, respectively), it notably improves forage yield and micronutrient accumulation, thus potentially delivering positive consequences at the farm level regarding economic performance and soil fertility when swards upon clayey soil types receive said urine in temperate agro-climatic regions (i.e., South West England in the current context). In contrast, faeces application in isolation hinders Se and Mn uptake, once again potentially delivering unintended consequences such as micronutrient deficiencies in areas of high faeces deposition. As it is unlikely that (b)ovine grazing fields will receive either urine or faeces in isolation, we also explored combined applications of both excreta types which demonstrates synergistic effects on N, Cu, and Zn uptake, with either synergistic or dilution effects being observed for P and S, depending largely on SOC levels. Additionally, interactions between excreta types can result in dilution or antagonistic effects on Mn and Se uptake. Notably, high SOC combined with faeces reduces Mn and Se in forages, raising concerns for grazed ruminant systems under certain biotic situations, e.g., due to insufficient soil Se levels typically observed in UK pastures for livestock growth. These findings underscore the importance of considering SOC and excreta nutritional composition when designing forage management to optimize nutrient uptake. It should be noted that these findings have potential ramifications for broader studies of sustainable agriculture through system-scale analyses, as the granularity of results reported herein elucidate gaps in knowledge which could affect, both positively and negatively, the interpretation of model-based environmental impact assessments of cattle and sheep production (e.g., in the case of increased yields [beneficial] or the requirement of additional synthetic supplementation [detrimental]).

Prevalence, antibiogram and molecular characterization of Listeria monocytogenes from ruminants and humans in New Valley and Beheira Governorates, Egypt

BackgroundListeriosis is a global health threat to both animals and humans, especially in developing countries. This study was designed to isolate Listeria monocytogenes from faeces; environmental samples; and cow, sheep and goat milk, as well as human stool, to study its molecular characteristics and antibiotic sensitivity in the New Valley and Beheira Governorates, Egypt. The isolation and identification of L. monocytogenes were carried out using traditional culture and biochemical methods, followed by antibiography, genus confirmation of some isolates and detection and sequencing of InlB genes via PCR.ResultsOut of 2097 examined samples, the prevalence of L. monocytogenes was 13.4% in animals; the prevalence was 9.2%, 2.4%, 25.4%, 4%, 42.4%, and 6.4% in cattle faeces, cattle milk, sheep faeces, sheep milk, goat faeces, and goat milk, respectively. However, the prevalence of L. monocytogenes was 8.3% in human samples. Both animal and human isolates showed 100% resistance to trimethoprim-sulfamethoxazole, and the isolates showed the highest sensitivity to flumequine (100%), amikacin (99.2%), gentamicin (97.6%), and levofloxacin (94.6%). Multidrug resistance (MDR) was detected in 86.9% of the tested isolates. The 16 S rRNA and inlB genes were detected in 100% of the randomly selected L. monocytogenes isolates. Phylogenetic analysis of three isolates based on the inlB gene showed 100% identity between faecal, milk and human stool isolates.ConclusionsFaeces and milk are major sources of listeriosis, and the high degree of genetic similarity between animal and human isolates suggests the possibility of zoonotic circulation. The high prevalence of MDR L. monocytogenes in both animal and human samples could negatively impact the success of prevention and treatments for animal and human diseases, thereby imposing serious risks to public health.

Molecular Detection of Eimeria bovis in Indonesian Beef Cattle Using Nested PCR Technique

Eimeria bovis is a pathogenic protozoan that causes cattle digestive tract infections, which can cause economic losses to farmers. It is necessary to develop specific and accurate detection methods to conserve livestock and prevent coccidiosis in Indonesia. This study aims to detect E. bovis by nested PCR and determine the relationship with reference sequences. A total of 167 samples of beef cattle feces were taken randomly from community farms spread across 18 provinces in Indonesia. The feces were examined natively, and then the oocysts were purified by the sugar flotation method, extracted by KIT extraction, and amplified by the nPCR technique. Positive samples were followed by sequencing and phylogenetic analysis using MEGA 11 software. This study used two pairs of primers (outer and inner) taken from ITS-1 molecular markers. As many as 96 out of 167 samples (57.5%) were positive for Eimeria spp., and 48 of the 96 samples were positive for Eimeria spp. (50%) were detected to be positive for E. bovis based on the presence of a 238 bp DNA fragment. The results of the phylogenetic analysis showed that the study sample formed a separate cluster from the E. bovis cluster from abroad. In conclusion, E. bovis was detected in 16 out of 18 provinces in this study, and the nPCR technique proved to have better sensitivity and specificity.

Comparative Study of Bacterial Microbiota Differences in the Rumen and Feces of Xinjiang Brown and Holstein Cattle

Xinjiang Brown cattle are a unique and widely distributed breed of dual-purpose cattle in the Xinjiang area of China, whose milk production performance differs from Holstein cattle. It has been known that variations in bacterial species of the gastrointestinal tract influence milk protein, fat, and lactose synthesis. However, the microbiota differences between Xinjiang Brown and Holstein cattle are less known. This study aims to compare the bacterial community composition of the rumen and feces of these two cattle breeds under the same dietary and management conditions. The 16s rRNA sequencing data and milk production of 18 Xinjiang Brown cows and 20 Holstein cows on the same farm were obtained for analysis. The results confirmed differences in milk production between Xinjiang Brown and Holstein cattle. Microbiota with different relative abundance between these two cattle breeds were identified, and their biological functions might be related to milk synthesis. This study increases the understanding of the differences in microbiota between Xinjiang Brown and Holstein cattle and might provide helpful information for microbiota composition optimization of these dairy cattle.

VARIATIONS OF COLIFORM BACTERIA ISOLATED FROM BALI CATTLE CALVES FECES, MARKETED AT BERINGKIT ANIMAL MARKET

Beringkit Animal Market is one of the largest animal markets in Bali. Which is a place for livestock transactions, especially bali cattle calves, which are usually used as breeds. Bali cattle calves need digestive tract microbes such as bacteria to carry out the digestives process. Coliform bacteria are one of the bacteria that play an important role in the digestive tract of bali cattle calves. Coliform bacteria are one of the bacteria that plays an important role in the digestive tract of bali cattle calves. Coliform bacteria which are included in the Enterobacteriaceae group consist of Escherichia coli, Enterobacter sp., Klebsiella sp., and Citrobacter sp. This research aims to determine what variations of Coliform bacteria are found in the feces of bali cattle calves marketed at the Beringkit Animal Market. This research was observational with the number of samples used were 50 fecal samples from 50 bali cattle calves. The samples were isolated on Eosin Methylene Blue Agar (EMBA) media and then tested and identified using the Indole Methyl Voges Proskauer Citrate (IMViC) principles and Blood agar test. The result showed that 82% of samples were positive Coliform. Based on the result, it can be concluded that in the feces of bali cattle calves marketed at the Pasar Hewan Beringkit contain Coliform bacteria consisting of Escherichia coli, Enterobacter sp., Klebsiella sp., and Citrobacter sp. with the highest percentage was Escherichia coli at 58%, followed by Citrobacter sp. at 12% and Enterobacter sp. and klebsiella sp. respectively 6%. It is necessary to count the total number of each bacteria to determine the pathogenicity of the bacteria.

PENGARUH MOLASES PADA EKOENZIM DAN FILTRAT CAMPURAN FESES SAPI POTONG DAN JERAMI PADI TERHADAP pH, TOTAL BAL DAN KADAR ALKOHOL

The beef cattle sector produces waste in the shape of cattle feces, which has yet to be effectively utilized. The farming sector produces waste in the shape of rice straw, but the processing has not been varied. The making of eco enzymes is one of the new ways to process beef cattle feces and rice straw into more useful. The research sought to examine how adding molasses affects the fermentation process of eco-enzymes derived from beef cattle feces and rice straw filtrate. The eco-enzymes were obtained from solid fermentation and liquid fermentation processes. Solid-state fermentation was conducted over a period of seven days, followed by the extraction procedure. The extracted filtrate was fermented facultatively anaerobic for 21 days. The research uses an experimental approach utilizing a completely randomized design or CRD there are 3 treatments, which is the addition of molasses at a dose of P1 = 2.5%, P2 = 5%, and P3 = 7.5%, each repeated 6 times. Data were processed using the ANOVA test and using Tukey’s test for further analysis. The results showed that molasses dosing had a significant effect (P&lt;0.05) on reducing the pH value to 3.34 - 3.68, producing total Lactic Acid Bacteria (LAB) 6.76x105 - 14.13x 105 CFU/ml, and alcohol content 6.12 - 13.69%.

Analysis of benzimidazole anthelmintic resistance in parasitic nematodes &lt;i&gt;Haemonchus contortus&lt;/i&gt; using nested isothermal amplification (PCR)

The purpose of the research is to monitor farms located in the European part of the Russian Federation to identify resistance to effects of benzimidazole anthelmintics in populations of nematodes Haemonchus contortus dwelling in the gastrointestinal tract of small cattle.Materials and methods. The studies were conducted in slaughterhouses located in the Moscow Region in 2023–2024. At the first stage, taxonomic identification of parasitic nematodes and larvae (L3) was made, and Strongylata species was determined from sheep. The study material was the abomasum with duodenum fragments and a distal rectum fragment with feces. For molecular studies, we used mature nematodes and H. contortus L3 larvae isolated from the abomasum and feces of small cattle brought to slaughterhouses in the Moscow Region from 8 regions of the European part of the Russian Federation: Moscow, Astrakhan, Oryol, Lipetsk, Tula, Bryansk regions, Stavropol and Dagestan. The studies were conducted at the premises of the Laboratory of Molecular Biology, the VNIIP – FSC VIEV. Statistical processing of the obtained data was made, and mean infection rates of parasitic nematodes (infection intensity and prevalence) were determined. Fifty-six DNA samples of nematodes H. contortus were examined using nested isothermal amplification (PCR) to identify gene alleles that determine resistance to benzimidazole drugs.Results and discussion. Molecular genetic studies of H. contortus DNA sampled from sheep brought from different Regions only detected homozygous individuals (100%) resistant to benzimidazole in the parasitic nematode population from the Oryol Region. Other regions identified only homozygous and heterozygous individuals susceptible to benzimidazole.