Cattle Fecal Samples Research Articles

Molecular serotyping of Shiga toxin-producing Escherichia coli (STEC) of animal origin in Iran reveals the presence of important non-O157 seropathotypes.

The present study reported the first serotyping (O:H typing) data documented in Shiga toxin-producing Escherichia coli (STEC) strains of animal origin in Iran in isolates recovered between 2008 to 2016. A total number of 75 STEC strains previously isolated from fecal samples of cattle, sheep, goats, pigeons, humans, and deer were assessed by different polymerase chain reaction (PCR) assays detecting the major virulence genes of STEC and phylogroups. Then, the strains were tested for the 16 important O-groups by PCR. Finally, twenty strains were selected for H-genotyping by PCR plus sequencing. The predominant serogroup was O113 which was detected in nine isolates (five cattle, 55.50%; two goats, 22.20%; two red deer, 22.20%) followed by O26 (3/3, 100%) in cattle, O111 (3/3, 100%) in cattle, O5 (3/3, 100%) in sheep, O63 (1/1, 100%) in pigeon, O75 (2/2, 100%) in pigeons, and O128 in goats (2/3, 66.60%) and pigeon (1/3, 33.30%). The most important recognized serotypes were O113:H21 in cattle (2/3) and goat (1/3), O113:H4 in red deer (1/1), O111:H8 in calves (2/2), O26:H11 in calve (1/1), O128:H2 in goats (2/3) and pigeon (1/3), and O5:H19 in sheep (3/3). One cattle strain carrying stx1, stx2, eae, and Ehly genes belonged to O26:H29 serotype. Most strains with determined O-groups were from the bovine source that highlighted the importance of cattle as reservoirs of potentially pathogenic serovars. The present study suggested that the top seven non-O157 serogroups should be assessed along with O157 in all future research and clinical diagnostics of STEC in Iran.

Assessment of Cryptosporidium Burden in Cattle in Federal Capital Territory, Nigeria

This study was undertaken to undertaken to assess the Cryptosporidium burden in cattle in the Federal Capital Territory (FCT), Nigeria. A total of 400 cattle fecal samples were screened for Cryptosporidium oocysts using Safranin-Methylene Blue staining technique. A questionnaire was used to analyze the possible factors driving the prevalence of Cryptosporidium infection in cattle in the study area. The overall prevalence of Cryptosporidium infection was 17.8% (71/400). The prevalence of Cryptosporidium in cattle was highest (32.4%, 22/68) in the Gwagwalada Area Council and lowest (12.1%, 8/66) in AMAC. There was a significant difference in prevalence of Cryptosporidium in cattle in the Area Councils (P= 0.010 < 0.005). There was a statistical difference in prevalence of Cryptosporidium and age, consistency of feces, breed, management practices and size of herd of cattle (P < 0.05) in the study area. There was no significant difference in sex and prevalence of Cryptosporidium infection in cattle in the study area (P > 0.05). This study demonstrated a widespread prevalence of Cryptosporidium infection in cattle in Federal Capital Territory. More worrisome is their prevalence in asymptomatic cattle as reservoirs that may sustain zoonotic cycle of transmission in the humans.

Prevalence of gastrointestinal parasitic infection in cows and buffaloes in Lower Dir, Khyber Pakhtunkhwa, Pakistan.

Gastrointestinal (GI) Parasitic infection is a hot issue for cattle management. There is variation of GI parasites effects in sex, age of cattle, drinking water condition, nutrition, and severity of infection. Studies on prevalence of GI parasites among cattle population in Dir Lower are lacking. A total of 40 farms were selected randomly in six tehsil namely Tehsil Adenzai, Tehsil Timergara, Tehsil Balambat, Tehsil Munda ,Tehsil Lalqala, Tehsil Khall. Freshly cattle fecal samples were collected randomly from the selected farms during March 2018 till December 2018. Out of 314 buffaloes and cattle examined 58.59% (184/314) were positive for eggs, cyst/oocyst of one or more species of GI Parasites. The prevalence of parasitic infection was higher in Buffaloes 63.55% (75/118) as compared to Cow 55.61% (109/196) but the difference was not significant (p>0.05) Entamoeba,spp, Moniezia spp, Haemonchus spp and Coccidian spp were found in this study. The non-treated animals indicated the highest percentage of infection in cow 57.71% (101/175) and buffalo 68.13% (62/91).GI parasite prevalence in female animal were higher female cow 62.58% (87/139) and female buffalo 77.33% (58/75) as compared to male. But the difference is non-significant (p> 0.05) Yearling calves had the lower rate of GI parasitic infection than adults. Future investigations are necessary to evaluate the economic loss due to GI parasites in cattle's.

Antimicrobial Resistance Profiles and Molecular Characteristics of Extended-Spectrum β-Lactamase-Producing Escherichia coli Isolated from Healthy Cattle and Pigs in Korea.

Antimicrobial-resistant bacteria isolated from food animals pose a major health threat to the public on this planet. This study aimed to determine the susceptibility profiles of Escherichia coli isolated from cattle and pig fecal samples and investigate the molecular characteristics of extended-spectrum β-lactamase (ESBL)-producing E. coli using gene identification, conjugation, and Southern blot approach. Overall 293 E. coli were recovered from cattle (120 isolates) and pigs (173 isolates) in 7 provinces of Korea during 2017-2018. Ampicillin, chloramphenicol, streptomycin, and sulfisoxazole resistance rates were the highest in pigs' isolates (>60%, p ≤ 0.001) compared to that in cattle (3-39%). Multidrug resistance (MDR) was higher in pig isolates (73%) than in cattle (31%), and the MDR profile usually includes streptomycin, sulfisoxazole, and tetracycline. Resistance to critically important antimicrobials such as ceftiofur, colistin, and ciprofloxacin was higher in weaners than those from finishers in pigs. The qnrS gene was detected in 13% of the pig isolates. Eight isolates from pigs and one isolate from cattle were identified as ESBL-producers and ESBL genes belonged to blaCTX-M-55 (n = 4), blaCTX-M-14 (n = 3), and blaCTX-M-65 (n = 2). Notably, the blaCTX-M-65 and qnrS1 genes were found to be carried together in an identical plasmid (IncHI2) in two isolates from finisher pigs. The blaCTX-M-carrying isolates belonged to phylogenetic groups B1 (n = 4), B2 (n = 2), A (n = 2), and D (n = 1). The blaCTX-M genes and non-β-lactam resistance traits were transferred to the E. coli J53 recipient from seven blaCTX-M-positive strains isolated from pigs. The blaCTX-M genes belonged to the IncI1α, IncFII, and IncHI2 plasmids and are also associated with the ISEcp1, IS26, IS903, and orf477 elements. These findings suggested the possibility of blaCTX-M-carrying E. coli transmission to humans through direct contact with cattle and pigs or contamination of food products.

A Nested-PCR assay for detection of Cryptosporidium spp. in cattle in Sulawesi Island, Indonesia

Cryptosporidium spp. is a protozoan parasite that cause cryptosporidiosis, a gastroenteric disease in wide range of animals and humans. Cryptosporidiosis affects significant economic loss in livestock production due to poor management practices, leading to faecal contamination. The parasite can be transmitted by ingestion of oocysts which have high survival ability in various type of environment and resistant to water chlorination treatment. In this study, we aimed to investigate the presence of Cryptosporidium spp. in various faecal samples of cattle (n=24, aged 1-2 years old) obtained from several farms in Sulawesi. Following faecal DNA extraction, we performed a nested-PCR assay using validated-Cryptosporidium 18S ribosomal RNA (18S rRNA) primers, resulting a size of amplified DNA approximately 770 bps. A total four samples (16.7%) were regarded as positive for Cryptosporidium spp., including samples obtained from farms in South Sulawesi (n=2), Central Sulawesi (n=1) and Gorontalo (n=1). This result confirms the risk of cryptosporidiosis not only among cattle in Sulawesi but also potential transfer of this parasite to humans especially from contaminated water and food. Therefore, early detection of the spread of oocysts is also crucial for monitoring animal health including cattle as production animals, human health, and environment.

Escherichia coli in different animal feces: phylotypes and virulence genes.

In this study, it was aimed to determine the phylogroups of Escherichia coli isolates from horse, cat, dog, sheep, cattle, and chicken feces samples and to investigate some important virulence genes of the isolates. For this purpose, a total of 600 feces samples, 100 from each animal species, were used as material. For the isolation of E.coli, feces samples were directly inoculated on MacConkey agar. The identification of the isolates was performed via phenotypic tests and species-specific multiplex Polymerase Chain Reaction (mPCR) method. PCR methods were used to phylotype E.coli isolates and to investigate virulence genes (bfpA, eaeA, LT, ST, Stx1, and Stx2). Of the total 600 E.coli isolates recovered in this study, 120 (20%), 269 (44.8%), 58 (9.7%), 19 (3.2%), 35 (5.8%), 56 (9.3%), 31 (5.2%), and 12 (2%) were identified as phylogroup A, B1, B2, C, D, E, F, and Escherichia clade I, respectively. While the virulence gene was detected in 149 (24.8%) E.coli isolates, no virulence gene was detected in 451 (75.2%) isolates. According to the analysis results, the most determined virulence gene was Stx1, while the least determined virulence gene was LT. In conclusion, in this study, when both the animal species and the number of E.coli isolates examined are considered, the data obtained are of great importance in epidemiological terms. However, the detection of virulence genes in 13.5% among phylogroup A, B1, and C isolates with commensal characteristics suggest that these isolates may show pathogenic characteristics with the virulence genes they contain.

PREVALENCE OF Blastocystis sp. IN CATTLE, GOAT AND SHEEP REARED BY DIFFERENT FARM MANAGEMENT SYSTEMS IN PAHANG, MALAYSIA

Blastocystis sp. is a familiar parasite in the gastrointestinal tract causing infection in humans and animals. The purpose of this study is to evaluate the Blastocystis sp. prevalence in three sorts of livestock; cattle, goats, and sheep in Pahang, Malaysia, which are reared under two management systems; intensive and semi-intensive farm management system. About 92, 96, and 65 cattle, goat, and sheep fecal samples respectively were collected from different farms around Kuantan, Bera, and Pekan. The fecal samples were cultured in Jones’ medium supplemented with 10% heat-inactivated horse serum and incubated at 37 ºC for 2 weeks, then observed under light microscopy daily. The total prevalence of Blastocystis sp. was 29.34% in cattle (27/92), 29.16% in goats (28/96), and 43.07% in sheep (28/65). Supported the results of this study, Blastocystis sp. prevalence was higher in sheep and livestock reared by a semi-intensive farm management system (44.38%). However, further study could be done for Blastocystis sp. subtypes identification to determine its genetic diversity. Notwithstanding, this study has provided additional knowledge on the prevalence of each livestock reared in farms around Pahang that serve as important information in understanding host-parasite relationships, besides determining the best farm management system to be applied by farmers.

Rapid detection of bovine rotavirus a by isothermal reverse transcription recombinase polymerase amplification assays

BackgroundBovine rotavirus A (BRVA) is considered to be the most common pathogen of severe diarrhea in cattle worldwide, which could lead to the death of newborn calves and cause the significant economic losses to the cattle industry. As a novel isothermal nucleic acid amplification technique, recombinase polymerase amplification (RPA) has been applied widely for the rapid detection of different important pathogens in human and animals.ResultsAn RT-RPA assay based on the real time fluorescence monitoring (real-time RT-RPA) and an RT-RPA assay combined with a lateral flow strip (LFS RT-RPA) were successfully developed by targeting the VP6 gene of BRVA. The RT-RPA assays allowed the exponential amplification of the target fragment in 20 min. After incubation of the LFS RT-RPA on a metal bath at 40 °C, the results were displayed on the lateral flow strip within 5 min, while real-time RT-RPA allowed the real-time observation of the results in Genie III at 42 °C. Both of the two assays showed high specificity for BRVA without any cross-reaction with the other tested pathogens causing diarrhea in cattle. With the standard RNA of BRVA serving as a template, the limit of detection for real-time RT-RPA and LFS RT-RPA were 1.4 × 102 copies per reaction and 1.4 × 101 copies per reaction, respectively. In the 134 fecal samples collected from cattle with diarrhea, the BRVA positive rate were 45.52% (61/134) and 46.27% (62/134) in real-time RT-RPA and LFS RT-RPA, respectively. Compared to a previously published real-time PCR, the real-time RT-RPA and LFS RT-RPA showed a diagnostic specificity of 100%, diagnostic sensitivity of 98.39% and 100%, and a kappa coefficient of 0.985 and 1.0, respectively.ConclusionsIn this study, BRVA was successfully detected in cattle fecal samples by the developed real-time RT-RPA and LFS RT-RPA assays. The developed RT-RPA assays had great potential for the rapid detection of BRVA in under-equipped diagnostic laboratory and the point-of-need diagnosis at quarantine stations and farms, which is of great importance to control BRVA-associated diarrhea in cattle herds.

Performance of diagnostic assays used to detect Cryptosporidium oocysts in faecal samples of cattle in Kuwait and genotyping of Cryptosporidium species

BackgroudCryptosporidium species are zoonotic protozoan parasites responsible for gastroenteritis in various animals and humans. The diagnosis of Cryptosporidium presents many challenges. This research attempted to match the diagnostic efficiency of the modified Ziehl–Neelsen technique (mZN), immunochromatographic assays (IC), and enzyme-linked immunosorbent assay (ELISA) for the detection of Cryptosporidium in faecal samples of cattle in Kuwait. In addition, polymerase chain reaction (PCR) was utilised to determine the predominant species infecting cattle in Kuwait and correlating the detected species with the results of different diagnostic tests used, the presence or absence of clinical signs, and the age group of the infected cattle.ResultsOf 400 analysed faecal samples, Cryptosporidium positive samples were 23%, 15.25%, and 14% using IC, ELISA, and mZN. IC had the highest sensitivity (74.07%), and mZN had the highest specificity (98.29%) using a composite reference standard (CRS) as a gold standard. The rapid IC test results in high false-positive results of cryptosporidiosis, whereas using mZN alone is insufficient to declare a negative faecal sample. Only 74.5% (35/47) of Cryptosporidium-positive samples by the three assays could be amplified by PCR. This study was the first to genotype Cryptosporidium in Kuwait. Cryptosporidium parvum (n = 26) was the dominant species detected from cattle samples, followed by C. andersoni (n = 6), C. bovis (n = 2), and C. raynae (n = 1). The findings showed a statistically relevant relationship between diarrhoea and the detection of Cryptosporidium spp. in faecal samples of cattle (p-value = 0.0003). Pre-weaned calves were the most vulnerable age group to Cryptosporidium spp. infection (p-value = 0.0007).ConclusionFor screening of Cryptosporidium infection in faecal samples, antigen detection or PCR methods combined with one of the microscopy techniques should be used. Cryptosporidium parvum was the prepoderant Cryptosporidium spp. recovered from cattle samples in Kuwait followed by C. andersoni. Cryptosporidium parvum is a significant risk factor for diarrhoea in pre-weaned calves. However, further study is needed as many other causes of diarrhoea in calves must be ruled out before a diagnosis of Cryptosporidium diarrhoea can be made.

Incidence and molecular characterization of ESBL-producing and colistin-resistant Escherichia coli isolates recovered from healthy food-producing animals in Pakistan

To investigate the occurrence and molecular features of ESBL-producing and colistin-resistant Escherichia coli isolates recovered from healthy food-producing animals in Pakistan. A total of 153 E. coli isolates were recovered from 250 faecal samples collected from livestock and poultry. The antibiotic susceptibility, resistant determinants and mobile genetic elements were determined for all the isolates. The clonal relatedness was analysed by MLST. Plasmids harbouring, localization and transferability of mcr-1 gene were carried out by Southern hybridization, S1-PFGE and transconjugation. Out of 153 E. coli strains, 49.01% isolates were ESBLs producers, whereas 18.95% were resistant to colistin and 84.31% of the isolates. Multidrug resistance was found in 84% of the isolates. The ESBL-producing E. coli in buffaloes, cattle, sheep, goat and broilers faecal samples were 60%, 74%, 54%, 50% and 68%, respectively. Among the ESBLs genes, blaCTX-M was the most prevalent group detected in 98.66%, while only mcr-1 of the colistin-resistant genes could be PCR amplified in 29 isolates. The common MGEs found were ISECP1 (35.13%), ISCR1 (33.78%), ISApl1 (20.27%) and Inti1 (58.10%). The most predominant Inc. types found were IncFIB 46.66%, followed by IncFIA 30.66%, IncFIC 26.66%, IncFrepB 26.66%, IncHI2 26.66%, IncP 22.66% and IncX4 21.33%. The most frequent sequence type detected was ST58. Southern blot and S1-PFGE confirmed the plasmid harbouring of mcr-1 gene. The co-occurrence of mcr-1 and ESBLs-encoding genes, along with MGEs in E. coli from healthy food animals in Pakistan, is a major concern. Antimicrobial resistance can be transferred from animals to humans by direct contact or via the food chain and environment. The prevalence and co-occurrence of ESBL and colistin resistance genes from food-producing animals is rare in Pakistan. To our knowledge, this is the first report to find ESBLs and mcr-1-harbouring E. coli from the faecal samples of the healthy food-producing animals in Pakistan. The presence of ARGs in association with MGEs, co-harbouring the virulence factors, as determined in the current study, is a severe threat to livestock and the human community as it has horizontally and food web transferability.

Application of Fourier transform infrared spectroscopy (FTIR) techniques in the mid-IR (MIR) and near-IR (NIR) spectroscopy to determine n-alkane and long-chain alcohol contents in plant species and faecal samples

n-Alkanes and long-chain alcohols (LCOH) have been used as faecal markers to assess the feeding behaviour of both wild and domestic herbivore species. However, their chemical analysis is time-consuming and expensive, making it necessary to develop more expeditious methodologies to evaluate concentrations of these markers. This work aimed to evaluate the use of Fourier Transform Infrared Spectroscopy (FTIR) technology in the near infrared (NIR) and mid infrared (MIR) intervals, for the determination of n-alkane and LCOH concentrations of different plant species and faecal samples of domestic herbivores. Spectra of 33 feed samples, namely L. perenne, T. repens, U. gallii, short heathers (mixture of Erica spp. and Calluna vulgaris), improved pasture grasses (mixture of L. perenne and A. capillaris), heath grasses (mixture of P. longifolium and A. curtissii), improved pasture species (mixture of L. perenne, T. repens and A. capillaris) and herbaceous species (mixture of all herbaceous species found in the plot)) and 181 faecal samples (cattle and horses) were recorded. In order to develop calibrations for the prediction of n-alkanes and LCOH concentrations, partial least squares (PLS) regression was used. Regarding the models developed for plant species, the best results were observed for the calibrations using NIR. The best external validation coefficients of determination (R2v) obtained were 0.90 and 0.79 for LCOH and n-alkanes, respectively. For faecal samples, in the NIR interval, results indicate similar external validation predictions (R2v) for both animal species (0.64). On the contrary, in the MIR interval, differences between cattle (0.70) and horses (0.57) faecal samples in R2v were observed. Regarding the models created for both animal species faeces, LCOH (C26-OH and C30-OH concentrations ranging from 713.3 to 4451.9 mg/kg DM, respectively; R2v values ranging from 0.72 to 0.95) and n-alkanes (C31 and C33 concentrations ranging from 112.8 to 643.2 mg/kg DM, respectively; R2v values ranging from 0.19 to 0.90) present in higher concentrations tended to be those with better estimates. Results obtained suggest that the selection of the technique to be used may depend on the type of matrix, being the homogeneity of the matrices one of the most important factors for its success. In order to improve the accuracy and robustness of the models created for the estimation of the concentrations of these markers using these methodologies, the database (greater variability) used for the calibrations of these models must be increased.

In vivo assessment of antibacterial activity of Cassia sieberiana stem bark extracts on enterohaemorrhagic Escherichia coli infection in Wister rats

Background: The acceptance of traditional medicine as an alternative form of health care has led researchers to further investigate the antimicrobial and other health benefits of medicinal plants including Cassia sieberiana. The objective of this study is to assess the in vivo antibacterial effects of C. sieberiana stem bark extracts on infections caused by human and animal isolates of enterohaemorrhagic Escherichia coli (EHEC) in Wister rats.&#x0D; Methodology: This in vivo study was designed for 21 days in 3 phases of 7 days each; adaptation, infection and treatment. Escherichia coli were isolated from aerobic cultures of human and cattle faecal samples and EHEC 0157 identified by serological typing using latex agglutination method. Aqueous and ethanol extracts of authenticated C. sieberiana stem bark, were prepared using standard method. Forty-five Wister rats were randomly divided into 9 groups (A-I) of 5 rats each. Rats in group A (uninfected with human or animal EHEC isolate and untreated) served as negative control while rats in group B (infected with EHEC animal isolate and untreated) and group C (infected with EHEC human isolate and untreated) served as positive controls. Rats in group C through group I were experimental groups that were either infected with human or animal EHEC isolate and treated, or uninfected but treated with ethanol and aqueous extracts of C. sieberiana. During each of the study phase, faecal samples were collected from the rats and processed for evaluation of EHEC count and to determine faecal occult blood. Data were analyzed using the Statistical Package for Social Sciences, version 20.0 and categorical variables were compared with Pearson’s Chi-square, with significant value taken as p&lt;0.05&#x0D; Results: Three EHEC isolates (2 from cattle and 1 from human) were identified from 22 E. coli isolates cultured from cattle and human faecal samples. Rats in group A (negative control) and those in group G (infected with EHEC human isolate and treated with ethanol extract), group H (not infected but treated with aqueous extract) and group I (not infected but treated with ethanol extract) were faecal occult blood negative throughout the study period. Rats in group B (infected with animal isolate of EHEC without treatment) were occult blood negative after infection on day 14 but positive on day 21, while rats in group C (infected with human isolate of EHEC without treatment) were occult blood positive on day 14 but negative on day 21. Rats in groups D, E and F infected with human and cattle EHEC isolates and treated, were faecal occult blood positive on day 14 but negative on day 21, with high colony counts recorded, cleared within 7 days of treatment by both aqueous and ethanolic extracts of C. sieberiana. &#x0D; Conclusion: The findings of this study confirmed the antibacterial potentials of C. sieberiana stem bark against EHEC. The beneficial effects of this plant extract should be exploited for commercial medicinal purposes.

Presence of Clostridioides difficile in cattle feces, carcasses, and slaughterhouses: Molecular characterization and antibacterial susceptibility of the recovered isolates

The aims of this study were to isolate and identify Clostridioides difficile from cattle feces and carcasses, and slaughterhouse samples, and to determine the molecular characteristics and antibacterial susceptibility of the recovered isolates. A total of 220 samples, including 100 cattle fecal samples, 100 cattle carcass surface samples, and 20 slaughterhouse samples were used as the study material. In total, 12 (5.45%) samples, including 11 (11%) cattle fecal samples and 1 (5%) slaughterhouse sample, were found to be positive for C. difficile. On the other hand, all of the carcass samples were negative for C. difficile. A total of 11 (91.66%) isolates, including 10 fecal isolates and 1 slaughterhouse wastewater isolate, were found to be positive for the presence of the toxin genes tcdA and tcdB, whilst 1 fecal isolate was found to be negative for both genes. In addition, 3 different ERIC-PCR profiles were identified in the 11 fecal isolates. The ERIC-PCR profile of the slaughterhouse wastewater isolate was found to be similar to one of the ERIC-PCR profiles obtained from the fecal isolates. All of the isolates were resistant to ciprofloxacin and levofloxacin. Considering that the agent is a spore-forming bacterium shed in feces, the detection of C. difficile isolates of different genotypes, some carrying toxin genes, suggests that feces and slaughterhouse wastewater carrying this bacterium may pose a risk for the contamination of carcasses. The current study revealed that hygiene conditions should be performed to the maximum extent in slaughterhouses.

Multidrug-Resistant Klebsiella variicola Isolated in the Urine of Healthy Bovine Heifers, a Potential Risk as an Emerging Human Pathogen.

Klebsiella variicola, a member of Klebsiella pneumoniae complex, is found to infect plants, insects, and animals and is considered an emerging pathogen in humans. While antibiotic resistance is often prevalent among K. variicola isolates from humans, this has not been thoroughly investigated in isolates from nonhuman sources. Prior evidence suggests that K. variicola can be transmitted between agricultural products as well as between animals, and the use of antibiotics in agriculture has increased antibiotic resistance in other emerging pathogens. Furthermore, in animals that contain K. variicola as a normal member of the rumen microbiota, the same bacteria can also cause infections, such as clinical mastitis in dairy cows. Here, we describe K. variicola UFMG-H9 and UFMG-H10, both isolated from the urine of healthy Gyr heifers. These two genomes represent the first isolates from the urine of cattle and exhibit greater similarity with strains from the human urinary tract than isolates from bovine fecal or milk samples. Unique to the UFMG-H9 genome is the presence of flagellar genes, the first such observation for K. variicola. Neither of the sampled animals had symptoms associated with K. variicola infection, even though genes associated with virulence and antibiotic resistance were identified in both strains. Both strains were resistant to amoxicillin, erythromycin, and vancomycin, and UFMG-H10 is resistant to fosfomycin. The observed resistances emphasize the concern regarding the emergence of this species as a human pathogen given its circulation in healthy livestock animals. IMPORTANCE Klebsiella variicola is an opportunistic pathogen in humans. It also has been associated with bovine mastitis, which can have significant economic effects. While numerous isolates have been sequenced from human infections, only 12 have been sequenced from cattle (fecal and milk samples) to date. Recently, we discovered the presence of K. variicola in the urine of two healthy heifers, the first identification of K. variicola in the bovine urinary tract and the first confirmed K. variicola isolate encoding for flagella-mediated motility. Here, we present the genome sequences and analysis of these isolates. The bovine urinary genomes are more similar to isolates from the human urinary tract than they are to other isolates from cattle, suggesting niche specialization. The presence of antibiotic resistance genes is concerning, as prior studies have found transmission between animals. These findings are important to understand the circulation of K. variicola in healthy livestock animals.

PREVALENCE OF HELMINTHIASIS IN CATTLE THROUGH FECES EXAMINATION IN MAGETAN REGENCY

This study aims to determine the prevalence of helminthiasis and find various kinds of egg worms in cattle through feces examination in Magetan Regency. A hundred cattle feces samples were collected from six animal markets in Magetan Regency. The Faecal examination was performed by native, sediments, and Flotation methods. The results of this study showed that there were 40 samples positively infected by a helminth, which indicated that the prevalence of helminthiasis in cattle was 40%. The kind of helminth were Oesophagostomum sp. (14%), Bunostomum sp. (8%), Mecistocirrus sp. (8%), Haemonchus sp. (4%), Toxocara vitulorum (3%), Trichuris sp. (2%), Strongyloides sp. (2%), and Fasciola sp. (11%).

The winner takes it all: dominance of Calicophoron daubneyi (Digenea: Paramphistomidae) among flukes in Central European beef cattle.

In Europe, paramphistomosis caused by Paramphistomum spp. was historically regarded as being of minor importance. However, Calicophoron daubneyi has recently been recognized as an emerging pathogen in Europe due to its increasing prevalence and negative impact on livestock production. In search for paramphistomid flukes, 5573 beef cattle fecal samples from 115 farms across the whole Czech Republic were examined from March 2019 to June 2021. The eggs of paramphistomid flukes were identified in 29.9% of samples. Internal transcribed spacer 2 sequences from 90 adult flukes and 125 fecal samples collected across Czech Republic confirmed C. daubneyi infection in the Czech beef cattle. Ninety mitochondrial DNA sequences obtained from adult C. daubneyi specimens revealed 13 individual haplotypes, two of them recorded for the first time. Although C. daubneyi is a new parasite in beef cattle herds in the Czechia, it clearly dominates the parasitological findings in the country's beef cattle. The common occurrence of C. daubneyi in most of the beef cattle herds indicates environmental conditions suitable also for the life cycle of Fasciola hepatica and risk of its emergence.

Recovery rates of Eimeria oocysts from cattle feces by Wisconsin sugar flotation method

The purpose of this study was to know the recovery rates of Eimeria oocysts through the course of flotation in sugar solution by WSFM. A fecal sample was collected from a calf infected with Eimeria bovis. Stage one of this study involved recovering oocysts from surface of sugar solution in a test tube with cover slips replaced at time points of 10, 20, 30, 40, 50, 60, 90 min, 2, 3, 4, 5, 6, 24, 48 hr., and 5 days. Oocysts were continuously detected up to 48 hr, with a linear increase in recovery counts for the first 2 hr and the average total count was 953 per gram. Recovery rates of oocysts were 8.1%, 13.3% and 16.4% at 10, 20 and 30 min, respectively. Stage 2 involved a set of 3 tubes. Cover slips were put at intervals of 10, 20 and 30 min on the first tube, while one cover slip was kept for 20 min on the second tube and one for 30 min on the third tube. No significant differences were detected in oocyst counts at 20 min between the first and second tubes, and at 30 min between the first and third tubes. Stage 3 involved the comparison of recovery rates at 10 min between sugar solutions with specific gravities of 1.200 and 1.266. There was no significant difference in oocyst counts between the two solutions. These results indicated that it takes time to recover all Eimeria oocysts from cattle fecal samples by WSFM, but total oocyst counts per gram can be estimated by counting oocysts detected at any sampling time points, such as 10, 20 or 30 min using one cover slip. No improvement of recovery rates was seen by using a higher concentrated sugar solution. Keywords: WSFM, Oocyst, Cattle

First isolation and genomic characterization of bovine parechovirus from faecal samples of cattle in Japan

A novel picornavirus was isolated from the faeces of a diarrhoeic cow using MA-104 cells at the third blind passage. This virus, named Den1/2021/JPN, was completely sequenced using total RNA from the cell culture supernatant by deep sequencing. The genome of Den1/2021/JPN had a standard picornavirus genome organisation with conserved picornaviral motifs. The 5' untranslated region harboured a type-II internal ribosomal entry site. Den1/2021/JPN was most closely related to a bovine parechovirus (Bo_ParV) named cow/2018/4, which has been recently identified in publicly available databases. Phylogenetic analyses and pairwise sequence comparison revealed that Den1/2021/JPN and Bo_ParV cow/2018/4 clustered with parechoviruses and were most closely related to Parechovirus E identified in birds of prey, exhibiting nucleotide sequence similarity of 64.2-64.5 %, 58.6-59.7 % and 66.3-66.4 % in the polyprotein, P1 and 2C+3 CD coding regions, respectively. This study presents the first report on the isolation of Bo_ParV. Den1/2021/JPN and Bo_ParV cow/2018/4, which are candidates for a novel species in the genus Parechovirus.

The diversity of Eimeria spp. in cattle in the Brazilian Semiarid region.

The aim of the present study was to find out the diversity of Eimeria species in cattle herds in the semiarid region of Brazil. Forty cattle fecal samples were collected from 20 farms in the Paraíba State, Northeast Brazil, and examined by centrifugation-floatation technique in sucrose solution. From each positive animal, 20 oocysts were photographed and measured. Infection by Eimeria spp. was detected in 17.12% (137/800) of the samples analyzed. All the farms had at least one animal that was positive for Eimeria spp. (100%; 20/20). In total, 2740 coccidia were photographed and measured. The species detected were: Eimeria bovis (35.1%); Eimeria canadensis (17.48%); Eimeria auburnensis (14.7%); Eimeria ellipsoidalis (9.7%); Eimeria zuernii (7.22%); Eimeria brasiliensis (4.56%); Eimeria bukidnonensis (3.97%); Eimeria illinoisensis (2.91%); Eimeria wyomingensis (1.42%); Eimeria alabamensis (1.27%); Eimeria cylindrica (0.76%); Eimeria pellita (0.54%); Eimeria ildefonsoi (0.21%); and Eimeria subspherica (0.07%). It was concluded that cattle in the semiarid region of Brazil were parasitized by 14 species of Eimeria. It is thinked that the sanitary management employed, as well as the system used for raising these animals, is the crucial point that leads to high rates of infection in this region.

Molecular detection of Campylobacter species from human and cattle faecal samples in Kilosa District, Tanzania

A growing number of Campylobacter species other than C. jejuni and C. coli have been considered as emerging human and animal pathogens but their contribution to human gastroenteritis is poorly documented. This study aimed at detecting Campylobacter species from human and cattle faecal samples in Kilosa District, Tanzania using molecular techniques without culture. Seventy (70) faecal samples were collected from five diarrheic and 65 non-diarrheic human patients attending Kilosa District Hospital in Tanzania from July to October 2019. During the same period, 30 faecal samples were also collected from healthy cattle in the same district. Genus and species identification of Campylobacter was conducted on the samples using molecular techniques [the polymerase chain reaction (PCR) and 16S rRNA sequencing]. Phylogenetic analysis was carried out by comparison of the 16S rRNA gene sequences to reference strains by the Neighbor-Joining method in MEGA X. Campylobacter species detection rate by PCR was 65.7% (46/70) and 20% (6/30) in humans and cattle, respectively. There were five human diarrheic cases, four of which were positive for Campylobacter and of these, two were children ≤15 years of age. In humans, 16S rRNA sequencing revealed that C. concisus was the most predominant species occurring at a frequency of 37.8% (14/37), followed by uncultured Campylobacter spp. 24.3% (9/37) and C. hominis 21.6% (8/37). The least represented species were C. jejuni and C. lanienae, all occurring at 2.7% (1/37). In cattle, five (100%) sequenced PCR products matched with C. lanienae. Phylogenetic analysis revealed that with the exception of C. lanienae, 16S rRNA sequences of Campylobacter species were closely related to the reference strains used (Percent identity: 90.51-96.56%). Based on our findings, we recommend that molecular techniques, mainly PCR be adopted for the direct detection of Campylobacter species during laboratory screening and surveillance studies.