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Comparison of results using electron microscope, immunodiffusion and fluorescent antibody analyses to detect rotavirus in diarrheic fecal samples of calves.

Seventy-nine diarrheic calf fecal samples were examined by electron microscopy, immunodiffusion and the fluorescent antibody technique for the presence of rotavirus (reovirus-like agent). Thirty-eight (48%) of the samples were positive by electron microscopy, 59% by immunodiffusion and 20% positive by fluorescent antibody technique analyses. Another 9% were suspect-positive by fluorescent antibody technique. Chymotrypsin treatment of the fecal samples increased the ease of observing the viral particles by electron microscopy and also intensified the immunodiffusion arcs obtained. Immunodiffusion analyses using specific antisera to the virus would appear to be a practical method of detecting rotavirus in diarrheic fecal samples.

Animals

Separation and properties of enterovirus and reovirus recovered from a fecal sample of calf with diarrhea.

In April, 1971, a disease with pyrexia and diarrhea as main symptoms broke out collectively among calves. Fecal samples were collected from calves involved and inoculated into bovine kidney (BK) cell cultures. As a result, the diarrheal feces of one calf were suspected to contain two agents simultaneously. One agent (C-121 E strain) was isolated from the primary infected BK cell culture fluid by terminal dilution passages. It had been predominant in replication and shown a cytopathic effect which gave rise to a granular appearance in the early stage of culture. The other agent (C-121 R strain) was isolated from the primary infected BK cell culture fluid by neutralizing the C-121 E strain contained in this fluid with antiserum against this strain. It caused cytoplasmic inclusion bodies to form. On the basis of their physico-chemical properties, the C-121 E strain was identified as bovine enterovirus and the C-121 R strain as reovirus. Serological tests indicated that some of the affected calves had been infected not only with the two strains isolated, but also with bovine viral diarrhea virus, bovine adenovirus type 7, and bovine parvovirus.

Animals

Detection of neonatal calf diarrhea virus, infant reovirus-like diarrhea virus, and a coronavirus using the fluorescent virus precipitin test.

Thirty-four calf and five infant fecal specimens were tested for the neonatal calf diarrhea virus (NCDV) and for the reovirus-like infantile diarrhea agent; respectively. The procedures used were the fluorescent virus precipitin test and immune electron microscopy. Fourteen of the calf stools contained detectable NCDV, and four of the five infant stools contained the reovirus-like human agent. Infectious NCDV was detected in four of the 34 calf fecal specimens when Madin-Darby bovine kidney cell cultures that had been inoculated with supernatant fluids from stool suspensions were stained with fluorescent antibody. The 20 calf stools that did not have detectable virus were examined for the bovine corona diarrhea virus. Coronavirus was found in two of these specimens.

Animals

Bacteriological quality of runoff water from pastureland.

Runoff from a cow-calf pasture in eastern Nebraska was monitored for total coliforms (TC), fecal coliforms (FC), and fecal streptococci (FS) during 1976, 1977, and 1978. Bacteriological counts in runoff from both grazed and ungrazed areas generally exceeded recommended water quality standards. The FC group was the best indicator group of the impact of grazing. Rainfall runoff from the grazed area contained 5 to 10 times more FC than runoff from the fenced, ungrazed area. There was little difference in TC counts between the two areas, but FS counts were higher in runoff from the ungrazed area and reflected the contributions from wildlife. Recommended bacteriological water quality standards, developed for point source inputs, may be inappropriate for characterizing nonpoint source pollution from pasture runoff. The FC/FS ratio in pasture runoff was useful in identifying the relative contributions of cattle and wildlife. Ratios below 0.05 were indicative of wildlife sources and ratios above 0.1 were characteristic of grazing cattle. Occasions when the FC/FS ratio of diluted cattle waste exceeded one resulted from differential aftergrowth and die-off between FC and FS. The FC/FS ratio and percentage of Streptococcus bovis in pasture runoff are useful indicators for evaluating the effectiveness of livestock management practices for minimizing bacterial contamination of surface water. The importance of choice of medium for the enumeration of FS in runoff derived from cattle wastes is discussed.

Enterococcus faecalis

Influence of chlortetracycline feeding on salmonellosis in calves. I. Rate and duration of shedding. II. Severity of illness.

Studies were conducted to test the influence of the feeding of chlortetracycline (CTC) on the fecal shedding of S. typhimurium subsequent to experimentally induced infection in calves. Levels of 0, 20, 50, and 100 gm CTC per ton of feed were fed to groups of calves for a two-week period before inoculation and the resulting level of shedding of S. typhimurium quantified. At the 50 gm/ton level, the feeding of CTC was associated with a significantly higher level of shedding than in non-CTC fed controls, and the duration of shedding was longer. Calves fed at 50 and 100 gm CTC/ton were affected much more severely by the inoculation than calves receiving no CTC. The same was true to a lesser extent in the calves fed 20 gm/ton. Observations made on each calf included changes in body temperature, time of onset, severity and duration of diarrhea, straining, and anorexia. Since the fecal output of salmonellae is increased at the level of 50 gm/ton, the authors conclude that this commonly used level of CTC feeding in calves contributes to the size of the salmonella reservoir in nature, thus increasing the risk of exposure to both man and animals alike and otherwise complicates the problems of salmonellosis.

Animal Feed

Reovirus-like agent (rotavirus) associated with neonatal calf gastroenteritis in France.

A reo-like virus (calf rotavirus) was shown to be associated with cases of neonatal calf diarrhea in France. The virus could be detected in more than 50 p. 100 of diarrheic fecal samples, while it was practically absent in control samples originating from healthy calves that had never had diarrhea. The results obtained by electron microscopy and immunofluorescent studies indicate that the virus is closely related or identical to the agent isolated originally in the United States.

Animals

Bovine salmonellosis: experimental production and characterization of the disease in calves, using oral challenge with Salmonella typhimurium.

A highly virulent strain of Salmonella tyhimurium was given orally to produce disease experimentally in 21 normal colostrum-fed calves 3 to 9 weeks old. The challenge inoculum varied from 10(4) to 10(11) organisms. The disease was characterized by fever, depressed attitude, and decreased appetite. Many calves given larger challenge dose levels also had diarrheic feces containing mucus, fibrin, and blood. Fecal cultures were positive for salmonella. Septicemia occurred in some calves (9 of 15 calves cultured were positive). Eleven calves died and 10 calves survived challenge exposure. Survival was inversely related to the size of the challenge inoculum and directly related (although to a lesser degree) to age of the calf. White blood cell total and differential counts were variable. Both neutropenia and neutrophilia were observed. Plasma proteins decreased markedly in calves with diarrhea, probably indicating fecal protein loss. Fibrinogen increased during the acute stages of diarrhea.

Administration, Oral

Whole-Genome Sequencing Reveals Co-Infection with Bovine Viral Diarrhea Virus, Bovine Enterovirus, and Caprine Parainfluenza Virus Type 3 in a Calf from a Cattle Herd in Xizang, China.

Although mixed viral infections are increasingly recognized as contributors to bovine diarrhea syndrome, diagnosing such co-infections remains challenging, particularly in high-altitude regions where surveillance is limited. In July 2024, a calf presenting with severe diarrhea and respiratory distress was identified on a cattle farm in Linzhi, Xizang, China. Using unbiased whole-genome sequencing (WGS) of the fecal sample, we assembled near-complete genomes of three distinct RNA viruses: two bovine viral diarrhea virus type 1 (BVDV-1) strains (subtypes 1v and 1q, designated BVDV-1/XZ87 and XZ87), one bovine enterovirus (genotype EV-E, designated BEV/XZ87), and one caprine parainfluenza virus type 3 (CPIV3/XZ87). The CPIV3/XZ87 genome exhibited 99.9% nucleotide identity to the goat-derived GS2017-2 strain from Jiangsu, China, raising the possibility of viral spread through livestock trade. Quantitative real-time PCR (RT-qPCR) confirmed the presence of all three pathogens (Ct values: 24.78 for BEV, 25.98 for CPIV3, and 31.28 for BVDV). This study provides the genomic evidence of a triple co-infection involving BVDV-1, BEV, and CPIV3 in Xizang. It illustrates the potential of WGS for unbiased pathogen detection in complex clinical specimens. The near-complete genomes generated here fill critical gaps in the virological surveillance of this epidemiologically under-sampled high-altitude region.

bovine enterovirus

Pathophysiologic changes due to coronavirus-induced diarrhea in the calf.

Extensive water, sodium, chloride, bicarbonate, and potassium losses occur in the diarrheal calf. The water loss is entirely from the extracellular space. In severe cases, hypovolemic shock occurs with the blood volume decreased by as much as one-half. Acidosis, which results from fecal bicarbonate loss, lactic acidosis, and renal dysfunction, results in tissue buffering, which in turn causes the efflux of cellular potassium ions. Although there is a total body potassium deficit, plasma potassium concentration is increased. This, in conjunction with an intracellular deficit, causes weakness, lethargy, and potassium cardiotoxicosis resulting in death. Hypoglycemia also contributes to the weakness and lethargy seem as calves become moribund. These losses from the body and shifts in fluids and electrolytes must be understood to develop the most effective rationale for supportive therapy.

Acidosis

Sarcocystis fusiformis infection in the coyote (Canis latrans).

Of 150 fecal samples from coyotes from northern Utah and southern Idaho, 21 contained coccidian sporocysts. These were found singly, contained four sporozoites and a granular residuum, and averaged 16.9 x 10.9 mum in size. None of four lambs inoculated with these sporocysts showed clinical signs of disease, and no parasites were found in their tissues. One calf inoculated with 135,000 sporocysts and another inoculated with 720,000 sporocysts eventually became unable to rise from recumbency. One was killed after 43 days; the other died at 50 days. Both calves had a mild mononuclear cell infiltration into the connective tissue adjacent to vessels in the heart, other striated muscles, kidney, and liver, and a moderate lymphoid hyperplasia in the spleen and lymph nodes. Schizonts were observed in a lymph node, and young sarcocysts were observed in skeletal and cardiac muscles. Four coyotes fed bovine heart infected with Sarcocystis passed sporulated sporocysts in their feces that were indistinguishable from those in the fecal samples from Utah and Idaho.

Animals

Rotavirus infections in adults in association with acute gastroenteritis.

During an epidemic of acute gastroenteritis in Helsinki, in March--May 1976, 18 out of 40 adult patients showed electron microscopic and/or serologic evidence for rotavirus infection. Rotavirus was most frequently seen in the fecal suspensions from 2 to 6 days after the onset of the symptoms but persisted in one patient for as long as 10 days. An increase in the complement-fixing (CF) serum antibody titers against the related Nebraska calf diarrhea virus (NCDV), or an initially high titer and subsequent significant decrease, was seen in all but one patient with rotavirus-positive feces, and in 6 additional patients. This suggests that using electron microscopy as the only diagnostic procedure a considerable number of rotavirus infections in adults remain undetected. Immune response against autologous or homologous rotavirus was also documented by immunoelectron microscopy. Complement-fixing antibody titers against NCDV decreased significantly from the convalescence values over a half-year observation period, but still remained clearly above the titers of a gastroenteritis-negative control population.

Acute Disease

Epidemiological and virological studies on outbreaks of acute gastroenteritis associated with rotavirus in primary schools in Osaka.

There have been three recent outbreaks of acute gastroenteritis in primary schools in Osaka prefecture caused by rotavirus: one in a school (TA) in April, 1974 and two in two other schools (TE and K) in May 1975. The morbidity from the disease was 2.4--15.8% for all age groups in the schools, and 20.1--34.1% for a certain age group. The disease lasted for 4 (K) to 14 days (TE). The first cases in schools TA and TE were followed by successive cases. The main clinical symptoms were higher frequencies of diarrhea (68--71.4%) and fever (78%) than in winter vomiting disease. There was no difference in the incidences of the disease in boys and girls. From observation on the disease in individual families, the incubation period seemed to be 3 days. Using paired sera and Neonatal Calf Diarrhea Virus (NCDV) as antigen, positive seroconversion was demonstrated by the complement fixation test and marked seroconversion by the indirect fluorescent antibody technique. With these techniques, specific antibody to NCDV was detectable in the sera from an early stage of illness. Rotavirus was found on electron microscopic examination of some fecal specimens of patients in TA, but not in those of patients in TE or K, although adenovirus was isolated from one patient. A serological survey of healthy children aged 0 to 12 showed that rotavirus is a common virus in Osaka.

Acute Disease

Distribution and seasonal prevalence of bovine lungworms in selected areas in western Montana.

In a survey (Oct, 1967, through Aug, 1969) of the distribution and seasonal prevalence of bovine lungworms in beef herds in 3 western Montana counties (Park, Gallatin, and Ravalli), infected cattle were detected in 27 of 35 herds. Results of 2,125 fecal examinations indicated that 7.8% of all cattle were infected with Dictyocaulus viviparus. Lungworms were found in 6.6% of 1,038 calves (less than 12 months old), 11.5% of 755 yearlings (1 to 2 years old), and 3.3% of 332 cows (more than 2 years old). A distinct seasonal pattern existed in calf infections, with initial passage of larvae beginning in late July or August and persisting until November or December. Exposure of calves in 1 portion of the study area apparently was delayed until November, with patent infections continuing through March. Spontaneous loss of infections normally occurred after 3 to 4 months of patency. The mean rate of larval excretion in the feces of all infected cattle was 0.37 larvae per gram of feces (l/g). Larval output was relatively uniform within each geographic area regardless of host age, but varied considerably from one portion of the study area to another. The potential role of wild ruminants (deer, elk, and moose) in the epizootiology of D viviparus is discussed with respect to western range utilization practices.

Animals

Coronaviral enteritis of young calves: virologic and pathologic findings in naturally occurring infections.

Coronavirus was detected in 113 (16.4%) of 689 calf enteric disease accessions received at the South Dakota Animal Disease Research and Diagnostic Laboratory from Mar 1, 1975 to Oct 31, 1976. During this period, modified live coronaviral vaccine was not in common use in this area. The virus was detected by negative stain electron microscopy of feces and intestinal contents and by fluorescent antibody examination of intestine and colon sections. Calves with coronaviral enteritis ranged in age from 1 day to 3 months, with most cases occurring between 1 and 7 days. At necropsy, a green, mucoid fecal cast was commonly seen in the colon. Histopathologic examination revealed necrotizing enterocolitis.

Animals

Immune electron microscopy of transmissible gastroenteritis virus and rotavirus (reovirus-like agent) of swine.

Immune electron microscopy (IEM) was developed as a diagnostic aid for detecting and identifying transmissible gastroenteritis virus and rotavirus (reovirus-like agent) in fecal and intestinal contents from cases of gastroenteritis in young pigs. Variables involved in use of direct IEM and its sensitivity were determined. Aggregates of virus coated with specific antibody were seen in virus samples mixed with homologous convalescent antiserum, but not in control samples containing preexposure serum or antibody directed against a heterologous virus. At least a ten fold enhancement of the sensitivity of direct IEM for virus detection was accomplished using indirect IEM employing rabbit anti-porcine IgG to further aggregate virus-antibody complexes. The technique was used to investigate the size and morphology of the porcine rotavirus. Particles ranged from 55 to 70 nm in diameter and had capsomere structures. Morphologically, the porcine rotavirus resembled the calf and human rotaviruses. By IEM, employing specific antiserums for each virus, porcine rotavirus was found to be antigenically related to these 2 viruses, but not to the reovirus type 3.

Animals