BACKGROUND: Diagnosis of Cryptosporidium species based on morphological characteristics is very limited and has no taxonomic value alone, and therefore the use of molecular methods removes these limitations to some extent. OBJECTIVES: The aim of this study was to determine the predominant Cryptosporidium genotypes in calves with diarrhea. METHODS: Study were conducted in calves aged less than 3 months for a period of 2 years. During the study period, 160 dung samples were collected from neonatal calves and examined first microscopically and then by molecular techniques. Stools were analyzed for the presence of Cryptosporidium oocysts by Sheather's Sugar Flotation Solution followed by Ziel-Neelsen staining method. DNA of parasite was extracted and multiplex nested-PCR protocol basis on the 18srRNA were done to identify three cattle-adapted species (C. andersoni, C. bovis and C. ryanae) plus the zoonotic species C. parvum. RESULTS: 110 fecal samples were collected from livestock in Alborz province and 50 fecal samples were collected from livestock in Shahroud city. Of the 160 animals examined, 90 were female and 70 were male. In total, out of 160 animals examined, 85 cases (53.12%) had diarrhea, of which 55 cases (34.37%) were positive using Ziel-Neelsen staining. Since all positive cases were related to diarrhea samples and related to calves under one month old, a significant relationship was observed between diarrhea status and the presence of this parasite (p < /em><0.05). In terms of seasonal distribution, no difference was observed in the rate of diarrhea and positive parasitic cases. The presence of 305 bp band in all Ziel-Neelsen positive samples confirmed the presence of C. parvum in all samples. CONCLUSIONS: Neonatal calves are more likely to be infected with Cryptosporidium parvum, as confirmed by the present study.

BACKGROUND: Diarrhea syndrome is associated with irrecoverable damages in the husbandry industry worldwide due to losses resulted from fatality, weight loss, growing weak calves and treatment costs. Hence, investigation of diarrhea causes in different areas is important to attempt management strategies to prevent and control it. OBJECTIVES: Present study was carried to investigate prevalence of some important entropathogens in diarrheic calves until three months old, in Shahrekord suburb husbandries. METHODS: Fecal samples were taken from 82 female calves in first day of diarrhea and were examined for isolation of salmonella, Escherichia coli, clostridium, cryptosporidium, and coccidia through common microbiological and parasitological methods. RESULTS: In general, prevalence of isolated organisms were: salmonella 36.6%, Escherichia coli 24.4%, clostridium 9.8%, cryptosporidium 9.8%, and coccidian 7.31%, and Escherichia coli K99 were isolated from four calves. The most prevalent pathogens were Escherichia coli and Salmonella. CONCLUSIONS: The calves are unavoidably exposed to infectious causes of diarrhea during their whole lifespan, because they acquire organisms from environment immediately after birth. Therefore, attempts at efficient management methods, hygienic principles and receiving enough colostrum, particularly in cold seasons, may be efficient in the control, prevention and decrease of diarrhea and its subsequent losses.

Due to increasing bacterial antibiotic resistance and the consumers’ tendency to choose organic products, cattle farmers are interested in alternative methods of calf diarrhoea treatment. This is a major challenge for veterinarians. Few methods of non-antibiotic treatment that bring satisfactory results have been reported in the related literature so far. In this article, the authors compare different non-antibiotic methods of diarrhoea prevention and treatment in calves. Among the alternatives discussed are herbs, probiotics, prebiotics and synbiotics, lactoferrin, and bacteriophages. It was found that the best results could be achieved through the use of pro-, pre- and synbiotics. However, the authors would like to point out that with the expansion of knowledge about the practical use of broad-scale bacteriophages, they could be the best alternative to antibiotics.

Fowl adenovirus can cause important diseases in chickens such as inclusion body hepatitis, hepatitis hydropericardium syndrome, and gizzard erosion and ulceration. Inclusion body hepatitis has been regularly reported from many countries. This is the first case report from Turkey, describing an outbreak of inclusion body hepatitis in broiler farms due to fowl adenovirus-8b (FAdV-8b). Broiler flocks with mortality about 10% were visited in Turkey, and necropsy was performed on dead birds. Samples were subjected to PCR assay to detect FAdV and other viral pathogens. After sequencing, phylogenetic analysis was performed and the nucleotide sequences of hexon genes were compared with the FAdV sequences data available in GenBank. Clinical signs such as anorexia, depression, ruffled feathers, huddling, and greenish diarrhoea were observed. Mortality started at the 8ᵗʰ day of age and ranged from 10% to 14%. Necropsy showed severe hepatitis, jaundice, and pancreatitis. The main necropsy findings included a pale, enlarged, haemorrhagic, and friable liver along with swollen and haemorrhagic kidneys and spleen. PCR and sequence analysis revealed the presence of fowl adenovirus serotype 8b (FAdV-E). This is the first report on characterisation and the pathological lesions associated with FAdV in broilers in Turkey. Our findings suggest that FAdV strains could be an emerging pathogen in Turkish broilers and could actively contribute to hepatitis and immunosuppression.

Porcine epidemic diarrhoea virus (PEDV) infection causes watery diarrhoea, vomiting, anorexia, and weight loss, especially among neonatal piglets, inflicting on them morbidity and mortality potentially reaching 90%–100%. Despite it being known that certain mammalian cell phases are arrested by PEDV, the mechanisms have not been elucidated, and PEDV pathogenesis is poorly understood. This study determined the effect of an epidemic PEDV strain on cell cycle progression. We observed the effect of the PEDV SHpd/2012 strain on an infected Vero cell cycle through flow cytometry and Western blot, investigating the interrelationships of cell-cycle arrest, the DNA damage–signalling pathway caused by PEDV and the phosphorylation levels of the key molecules Chk.2 and H2A.X involved upstream and downstream in this pathway. PEDV induced Vero cell-cycle arrest at the G1/G0 phase. The phosphorylation levels of Chk.2 and H2A.X increased with the prolongation of PEDV infection, and no significant cell-cycle arrest was observed after treatment with ATM or Chk.2 inhibitors. The proliferation of PEDV was also inhibited by treatment with ATM or Chk.2 inhibitors. PEDV-induced cell-cycle arrest is associated with activation of DNA damage–signalling pathways. Our findings elucidate the molecular basis of PEDV replication and provide evidence to support further evaluation of PEDV pathogenesis.

Enteropathogenic Escherichia coli (EPEC) is one of the main pathotypes causing gastroenteritis, particularly in young immunocompromised hosts. The study reports the prevalence, characterisation, and molecular epidemiology of EPEC from piglets in northeastern India. A total of 457 faecal samples were collected, from which 1,286 E. coli strains were isolated and screened by PCR. The resultant EPEC strains were serotyped and phenotypically characterised for resistance against 15 antimicrobials. Also, the phylogenetic sequence was analysed for 11 selected strains. A total of 42 strains (3.26%) belonged to atypical EPEC, of which, 15 (35.71%, and 2.29% of the 654 strains from this farm type) were isolated from organised and 27 (64.29%, and 4.27% of the 632 strains from this farm type) from unorganised farms; further, 5 (11.90% of the EPEC strains and 1.51% of the 330 strains from this breed) were isolated from the indigenous breeds and 37 (88.10%, and 3.87% of the 956 strains from this breed) from crossbred piglets. Serogroups O111 (11.9%) and O118 (7.14%) were the most prevalent of the 10 present. Sequence analysis of a length of the eaeA gene of 11 isolates of the region showed them to have 100% homology with each other and their identity ranged from 99.4% to 99.7% with GenBank reference sequences. All the EPEC isolates were multi-drug resistant, showing the highest resistance to amoxicillin (80.9%) and cephalexin (76.19%). The study highlighted the association of EPEC with piglet’s diarrhoea in northeastern India. EPEC isolates belonged to many serotypes and phenotypically all were multi-drug resistant with close genetic homology.

Canine parvovirus (CPV) disease is one of the most threatening to domestic and wild dogs. A total of 132 clinical samples were isolated from domestic dogs with diarrhoea from Henan, Hubei, Jiangsu, and Anhui provinces from 2016 to 2017, and 56 were positive for CPV-2 by PCR. A phylogenetic tree was constructed for the isolate sequences incorporating 53 non-Chinese reference strains. VP2 sequences showed the strains mainly to be new CPV-2a/2b and CPV-2c genotypes. The Ala5Gly, Phe267Tyr, Ser297Ala, Tyr324Ile, Gln370Arg, Asn426Asp or Asn426Glu, and Thr440Ala sites in the VP2 protein antigenic region were found to have high mutation rates. The VP2 tertiary structural model shows that the change at these mutation points is a factor for the changes in the protein structure. Significant differences between the Central Chinese strains and others were found, indicating that evolution is geographically related and extended in major regions. The homology between the identified strains confirmed their relationship. Phylogenetic analysis indicated that the common genotypes in the same clusters differ slightly in homology and evolutionary history. This epidemiological study enriches the available data and serves as an important reference for studies on the evolution of CPV and selection of vaccines in China.

OBJECTIVE To evaluate the usefulness of intestinal biomarkers in determining the presence of intestinal epithelial damage in neonatal calves with diarrhea caused by 4 etiologic agents. ANIMALS 40 neonatal calves that were healthy (n = 10) or had diarrhea (30). PROCEDURES The study was a cross-sectional study. Results of hematologic analyses and serum concentrations of intestinal fatty acid–binding protein (I-FABP), liver fatty acid–binding protein (L-FABP), trefoil factor 3 (TFF-3), Claudin-3 (CLDN-3), γ-enteric smooth muscle actin (ACTG2), intestinal alkaline phosphatase (IAP), interleukin-8 (IL-8), platelet-activating factor (PAF), and leptin (LP) were compared among calves grouped according to whether they were healthy (control group; G-1) or had diarrhea caused by K99 Escherichia coli (G-2; n = 10), bovine rota- or coronavirus (G-3; 5 each), or Cryptosporidium spp (G-4; 10). RESULTS Across the 3 time points at which blood samples were obtained and evaluated, the groups of calves with diarrhea generally had markedly higher mean serum concentrations of L-FABP, TFF-3, IAP, IL-8, and LP, compared with the control group. In addition, G-2 also consistently had markedly higher mean serum concentrations of I-FAB and ACTG2 and lower mean serum concentrations of CLDN-3, compared with the control group. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that degree of intestinal epithelial damage differed among calves grouped by the etiologic agent of diarrhea and that such damage might have been more severe in calves with diarrhea caused by K99 E coli. Additionally, our results indicated that serum concentrations of I-FABP, L-FABP, TFF-3, IAP, IL-8, ACTG2, LP, and CLDN-3 were useful biomarkers of intestinal epithelial damage in calves of the present study.

OBJECTIVE To investigate the activities of gelatinases (matrix metalloproteinase [MMP]-2 and MMP-9) and serine proteases in the colorectal mucosa of Miniature Dachshunds (MDs) with inflammatory colorectal polyps (ICRPs). ANIMALS 15 MDs with ICRPs and 5 dogs with non–ICRP-related large bowel diarrhea (controls). PROCEDURES Zymographic methods were used to evaluate the activities of MMP-2, MMP-9, latent forms of MMP-2 and MMP-9 (pro–MMP-2 and pro–MMP-9), and serine proteases in inflamed and noninflamed tissue samples from MDs with ICRPs and in noninflamed tissue samples from control dogs. The associations of serine protease activities with MMP-2 or MMP-9 activity were also analyzed. RESULTS Activities of pro–MMP-2 and pro–MMP-9 were detected in most tissue samples, regardless of the tissue type, whereas activities of MMP-2 and MMP-9 were not detected in control tissue samples. In the inflamed tissue samples from MDs with ICRPs, the activities of MMP-2, pro–MMP-9, and MMP-9 were significantly higher than those in the noninflamed tissue samples from those dogs. Serine protease activities were significantly higher in the inflamed and noninflamed tissue samples from MDs with ICRP, compared with findings for control tissue samples. A weak correlation was detected between serine protease activities and MMP-9 activity. CONCLUSIONS AND CLINICAL RELEVANCE Study results suggested that gelatinase and serine protease activities are upregulated in the colorectal mucosa of MDs with ICRPs, possibly contributing to the pathogenesis of this disease through the functions of these enzymes in degradation of extracellular matrix and promotion of inflammatory cell migration and inflammatory responses.