Fecal Samples Of Diarrheic Calves Research Articles

Detection of Cryptosporidium parvum in calf feces using microscopical, serological, and molecular methods

Cryptosporidiosis is a critical gastrointestinal disease in calves. This study examined 150 fecal samples of diarrheic calves collected from the eastern region of Saudi Arabia for detection of Cryptosporidium parvumusing the Modified Ziehl-Neelsen (MZN) method, Enzyme-Linked Immunosorbent Assay (ELISA), and conventional Polymerase Chain Reaction (PCR). The performance of these methods was assessed using diagnostic accuracy tests. The present study identified C. parvum oocysts in fecal samples by modified Ziehl-Neelsen 40/150; 26.66%, ELIZA 60/150; 40%, and PCR 78/150; 52% methods, respectively. The microscopic method revealed higher specificity 65.27% than the ELIZA 51.38%, while the ELIZA showed higher sensitivity 32.05% than MZN method 19.23%. However, MZN and ELIZA methods were unsatisfactory diagnostic tools compared with the PCR as the area under the curve values in Receiver Operator Characteristic (ROC) analysis were less than 0.6. Furthermore, using the kappa analysis test revealed no agreement between MZN and ELISA methods compared with PCR at P<0.05.

Isolation and whole protein characterization of species A and B bovine rotaviruses from Chinese calves.

Rotaviruses (RVs) account for severe diarrhea in children and young animals globally. In the current study, the fecal samples of diarrheic calves from a beef farm in Inner Mongolia were screened for RVA by ELISA and RT-PCR, followed by culture of three positive RVA samples in the MA-104 cell line. After 10 blind passages, cytopathic effects (CPE) appeared as detachment, granulation, and clustering of the inoculated cells. The virus isolates were identified by RT-PCR (VP6 gene RVA) and ESI-LC-MS/MS for whole protein sequencing. The protein sequences demonstrated the presence of two strains from species A rotavirus and one RVB strain; RVA/Cow-tc/CHN/35333/2019/G6P[5] was mixed with one RVB strain (RVB/Cow-tc/CHN/35334/2019/G5P[3]) in two samples, and RVA/Cow-tc/CHN/10927/2019/G8P[7] was found in one sample. They are of genotype constellations (G6-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3), (G8-P[7]-I5-R1-C1- M2-A1-N1-T1-E1-H1), and (G5-P[3]-I3-R5-C5-A5-N4-H5), respectively. Besides, phylogenetic analysis of the obtained sequences demonstrated viral evolution.

Etio-Epidemiological Studies on Diarrhoea in Cattle and Buffalo Calves

Calf diarrhoea is the most commonly encountered disease syndrome and significant cause of economic losses in dairy industry. Present investigation was undertaken to find out the prevalence of causative agents of diarrhoea in the bovine calves for a period of one year. The effect of age, sex, season and parity of dam was also studied. E. coli was the major organism (86.00 %) observed in the faecal samples of the diarrhoeic calves followed by rotavirus, Eimeria spp. and Amphistomes (15.00 % each); Toxocara spp. (12.00 %); Strongyles (9.00 %); Cryptosporidium spp. (6.00 %); Trichuris spp. (5.00 %); and Salmonella spp. and Strongyloides spp. (3.00 % each). The prevalence of rotavirus, Cryptosporidium spp. and Eimeria spp. was found significantly higher in buffalo calves and crossbred calves than cow calves and Gir / local non-descript calves, respectively. The prevalence of Toxocara spp., Amphistomes and Strongyles in diarrhoeic buffalo calves was significantly higher than cow calves. Highest prevalence of E. coli and rotavirus was observed in faecal samples of diarrhoeic calves of 0-15 days age group. Rotavirus was not detected in faecal samples of diarrhoeic calves above 60 days age. The susceptibility of bovine calves for E. coli and rotavirus was found decreased with the advancement of the age. The prevalence of Salmonella spp. in diarrhoeic faecal samples of bovine calves was observed only in 16-60 days age whereas Cryptosporidium spp. was found only in 0-30 days age. The most of the parasitic infestations were observed after 30 days of age in calves. The calves of both sexes were equally susceptible to different causative agents of diarrhoea. The prevalence of E. coli and most of the helminth ova in the faecal samples of diarrhoeic calves was found maximum during rainy season whereas the rotavirus was observed mostly during winter season. The prevalence of E. coli, Salmonella spp., rotavirus and Cryptosporidium spp. was found highest in the faecal samples of the diarrhoeic calves of first or second parity dams.

Detection and first molecular characterisation of three picornaviruses from diarrhoeic calves in Turkey.

The involvement of picornaviruses in calf diarrhoea was evaluated by the analysis of 127 faecal samples collected from diarrhoeic calves during 2014-2016. Virus detections were carried out by PCR using generic or specific primer pairs. One-third of the faecal samples (33.86%) were found to be positive for one or more of the studied viruses. Bovine kobuvirus was detected in 22.83%, bovine hungarovirus in 11.02%, while bovine enterovirus 1 in 5.51% of the samples. The sequences of the PCR products indicated the existence of novel variants in all the three virus species. When comparing the partial sequences, the nucleotide sequence identities between our newly detected viruses and those previously deposited to the GenBank ranged between 76 and 99%. Phylogenetic analyses revealed a novel lineage within the species Hunnivirus A. Our findings suggest that these viruses should be regarded as possible aetiological agents of calf diarrhoea. Based on the newly determined sequences, we designed and tested a new generic PCR primer set for the more reliable detection of bovine hungaroviruses. This is the first report on the molecular detection of the presence of bovine hungarovirus, bovine kobuvirus and bovine enterovirus 1 in the faecal samples of diarrhoeic calves in Turkey.

The prevalence of coronavirus in fecal samples of neonatal calf diarrhea using electron microscopic examination

BACKGROUND:Neonatal calf diarrhea (NCD), also known ascalf scours, is caused by viruses, bacteria, and parasites.Coronavirus is one of the important causes of NCD syndrome.Electron microcopy has been used for demonstration of virusesin fecal samples of diarrheic calves since 1969. OBJECTIVES:The aim of this study was to determine the prevalence ofcoronavirus in fecal samples of neonatal calf diarrhea usingelectron microcopy examination of fecal samples. METHODS:For the purpose of this study, a total of 100 cases (under 2 monthsof age) of diarrheic calves from 25 commercial farms in fourdistricts of East Azerbaijan province of Iran were used. FreshFecal samples using sterile swab were collected from every calfwith clinical diarrhea. All samples were transferred into a sterilecontainer and then were sent to the laboratory to be kept in -20oCfreezer until examination. Electron microscopic examinationwas applied to all fecal samples. Chi-square test was used toanalyze the data. RESULTS:According to the results of this study,coronavirus was present in 15% of cases and a significantdifference was found between the prevalence rate and theseasons (p<0.05). The highest prevalence was recorded in winter(28%) and the least prevalence was seen in summer (4%). Therewas not any significant difference in both age groups anddifferent districts according to the prevalence rate of infection.CONCLUSIONS: Neonatal calf diarrhea is a multi-etiologicalsyndrome. Among the viral diarrhea, bovine coronavirus is oneof the major causes of this syndrome. For detection ofcoronavirus in feces, electron microscope examination is a goldstandard diagnostic technique. It seems that coronavirus can beconsidered as one of the important etiologies of neonatal calfdiarrhea in east Azerbaijan of Iran.

Cryptosporidium spp., comparative diagnosis and geospatial distribution in diarrheic calves from dairy farms, Valdivia, Chile

ABSTRACTObjective. To determine the Cryptosporidium spp. infection frequency by using Ziehl-Neelsen and Auramine stains on samples obtained from diarrheic calves from milking farms of the Valdivia province. To compare both diagnostic tests and to determine the geospatial distribution of the infections caused by this protozoan. Materials and methods. 221 fecal samples of diarrheic calves of 24 milking farms of the Valdivia province were studied. The processing and analysis of the samples was done by Ziehl-Neelsen (ZN) and Auramine (AU) staining techniques, and the results were compared by McNemar statistical test and the concordance level was determined by kappa index. A map was also generated to determine the geospatial distribution of Cryptosporidium infections. Results. 57.9% of all the animals tested were classified as positive with the ZN stain test, while 55.6% of all the animals turned out positive for the AU stain test. The McNemar test showed no significant difference between both diagnostic techniques (p&gt;0.05), while the kappa index showed proper concordance between tests (κ=0.73). 100% of the farms studied showed protozoan presence demonstrating the broad distribution of the parasite, however, and considering the previous factor, it was not possible to determine geospatial associations for the parasite distribution. Conclusions. The infection frequency of Cryptosporidium is higher than 50% in the milking farms studied from the Valdivia province. No difference between the Ziehl-Neelsen and Auramine staining techniques was demonstrated showing very consistent results. It was possible to detect that the number of farms infected correspond to 100% of the farms analyzed.

Determination of G and P type diversity of group A rotaviruses in faecal samples of diarrhoeic calves in Kashmir, India

To determine the genetic diversity of group A rotaviruses in bovine calves in Kashmir, India. Of 200 diarrhoeic faecal samples collected from calves, aged between 0 and 6 months and screened by polyacrylamide gel electrophoresis (PAGE), 31 were detected positive for group A rotaviruses. On G and P genotyping by reverse transcriptase-polymerase chain reaction (RT-PCR), G10P[11] turned out to be predominant (80·64%) combination followed by G8P[11] (7·7%). One (3·84%) sample carried mixed infection of G8+G10P[11]. Two (7·7%) samples belonged to P[11] genotype, but their G genotype specificity could not be established. This study revealed the ambiguity in RT-PCR typing method. All the samples that turned out to be G10 by Isegawa et al. (1993; Mol Cell Probes7, 277) primers could be amplified by G3 specific primers of Gouvea et al. (1990; J Clin Microbiol32, 1338). However, on homology study of their VP7 gene sequence, the strains turned out to be G10. Rotavirus is prevalent in diarrhoeic calves in Kashmir, India, and G10P[11] is the predominant genotype in circulation. There is evidence of mixed infection. Even though RT-PCR method is the quick way to type the strains, there is need to generate more sequence data to improve the specificity of typing primers. Rotavirus is a significant cause of diarrhoea in calves. RT-PCR typing method needs to be supported by the sequence data, and there is need to re-evaluate the primers used for typing.

Yeasts in the gastrointestinal tract of preweaned calves and possible involvement of Candida glabrata in neonatal calf diarrhea

To examine the possibility of a mycotic involvement in neonatal calf diarrhea (NCD) the presence of fungi was assessed in (a) the intestinal contents of dead calves and fecal samples submitted for routine laboratory examination, (b) fecal specimens, sampled once in winter and once in summer, of calves raised on 2 farms with different management systems, and (c) mucosal scrapings of various segments of the digestive tract of a diarrheic calf, massively shedding Candida glabrata.C. glabrata was the most prevalent fungal species isolated from the routine samples. It was the only fungus which was shed by the calves on the 2 farms, for continuous, more or less prolonged periods, but exclusively in the winter months. Diarrhea and C. glabrata shedding seemed to be associated. C. glabrata colonized the abomasum (the functional equivalent of the monogastric stomach) but not the other segments of the digestive tract of the euthanized calfBased on the findings of this study it seems that while some yeast species may be considered as commensals of the digestive tract of calves, and consequently their isolation from intestinal contents or fecal samples has no clinical significance, others, such as C. glabrata may be involved in enteric pathogenic processes. Moreover, characteristics of the culture, previous chemotherapeutic treatments, the animal's age and possibly climatic conditions should be taken into account before deciding on the fungal isolate's clinical relevance. Determination of mycotic involvement in NCD by routine mycological examination of intestinal contents and fecal samples of diarrheic calves may be useful to avoid unnecessary and potentially harmful antibacterial therapy.