Presence of bacterial glycocalyx and fimbriae on Pasteurella haemolytica in feedlot cattle with pneumonic pasteurellosis.

This investigation was conducted to determine if Pasteurella haemolytica within feedlot cattle affected by pneumonic pasteurellosis express fimbriae (pili) and bacterial glycocalyx. Bacteriological culture of pulmonary tissue from three calves with fibrinous pneumonia resulted in heavy growth of P. haemolytica. Transmission electron microscopy of the lungs showed numerous microcolonies of gram-negative bacteria with morphology typical of Pasteurella haemolytica. The cells within these microcolonies possessed bacterial glycocalyces which stained with ruthenium red. Glycocalyx-encased microcolonies were also present in specimens examined by scanning electron microscopy. Typical P. haemolytica cells were evident in a tracheal specimen and these bacteria had radial glycocalyces consistent with polysaccharide and proteinaceous material condensed on linear structures suggestive of fimbriae. The pathogenetic importance of the bacterial glycocalyx and fimbriae in shipping fever pneumonia has yet to be established but their presence in clinical cases of Pasteurella pneumonia in feedlot cattle further supports a possible role in the initiation and progression of this disease as well as bacterial resistance to antimicrobial agents.

Electron microscopic examination of cells of Pasteurella haemolytica-A1 in experimentally infected cattle.

Several modern electron microscopy techniques were used to examine Pasteurella haemolytica (biotype A, serotype 1) (strain B122) recovered from experimentally infected cattle and in situ within the lung tissue of experimentally infected cattle. Glycocalyx four to five times thicker than that seen on P. haemolytica grown in vitro was evident on bacterial cells recovered from live infected calves by pulmonary lavage. Fimbriae were also present on cells recovered by lavage. A thick glycocalyx was also seen on P. haemolytica-A1 within the lungs of experimentally infected cattle at necropsy. In summary, cells of P. haemolytica-A1 in experimentally infected cattle have fimbriae and glycocalyx on their cell surfaces and these structures appear to be important in bacterial colonization of the bovine respiratory tract and pathogenesis of shipping fever (Pasteurella) pneumonia.

Electron microscopic description of glycocalyx and fimbriae on the surface of Pasteurella haemolytica-A1.

Several electron microscopic techniques were used to examine the surface of cells of Pasteurella haemolytica (biotype A, serotype 1) grown in vitro. All methods showed the presence of a very extensive glycocalyx on logarithmic phase (6 h) cells grown in liquid media. The anionic glycocalyx of these cells stained well with ruthenium red, but collapsed during dehydration for electron microscopy unless stabilized with specific antibodies. When the same techniques were used to examine cells in the stationary phase (18 h) the glycocalyx was much reduced. Large numbers of fimbriae were seen on both 6 h and 18 h cells grown in fluid media without shaking. In summary, logarithmic phase cells of P. haemolytica have both fimbriae and extensive anionic glycocalyx at their surface and we suggest that either or both of these structures may be important in the colonization of the bovine respiratory tract and the subsequent pathogenesis of Pasteurella pneumonia.

Enterotoxigenic colibacillosis in colostrum-fed calves: pathologic changes.

Enterotoxigenic colibacillosis was experimentally produced in 8 of 9 colostrum-fed calves orally given 10(11) Escherichia coli. The eight calves developed profuse diarrhea accompanied by dehydration and depression. At 12 hours after exposure, all calves were euthanatized for necropsy and for collection of tissues for microscopic examination. Histopathologic changes included stunted villi in the jejunum and ileum, focal degeneration and exfoliation of absorptive epithelial cells at the tips of jejunal and ileal villi, and focal emigration of neutrophils which was especially prominent above the dome area of aggregated lymphatic follicles (Peyer's patches). A layer of E coli adhered to the epithelial surface of the jejunum and ileum. In the duodenum, lesions were minimal or absent and bacteria were not adhering to the mucosa. Histopathologic changes were not observed in other tissues. In two calves examined 24 hours after they were inoculated and in two calves euthanatized 24 to 36 hours after spontaneously developing enteric colibacillosis, lesions were similar to those observed in the calves at 12 hours after exposure.

The prevention and control of epidemics of acute indifferentiated diarrhea of beef calves in Western Canada.

It is frequently evident that outbreaks of diarrhea occur in spite of apparent "good management" and "good calving conditions". This observation underlies the fact that we still do not understand many of the epidemiological factors which contribute to calf diarrhea outbreaks. For example, we still lack biological criteria by which to judge the degree of crowding and the degree of stress. Nevertheless, application of the principles described above will prevent or decrease the severity of many annual epidemics. To be successful, a program of prevention and control should be discussed with producers long before the calving season, preferably during the preceding summer or fall. Implementation of a complete program may take several calving seasons and producers should be made aware that prevention by improved management is an on-going, evolutionary process. More and improved vaccines are becoming available; however, as is the case with most biologicals, their impact cannot be determined until after they have been used for several years. They should be recognized as only one of several managent tools at the disposal of the veterinarian and livestock producer.

A comparison of dichromate solution floatation and fecal smears for diagnosis of cryptosporidiosis in calves.

Fecal samples from eleven calves experimentally infected with cryptosporidia were examined by two methods to evaluate their sensitivity and ease of use as diagnostic techniques. Comparison of a dichromate solution floatation and fecal smear techniques indicated that the former method was more sensitive. Oocysts were detected in feces earlier and for a longer period following exposure, and were easier to visualize using the dichromate solution floatation procedure. In addition, the dichromate solution floatation technique eliminated Candida albicans which can sometimes be confused with cryptosporidial oocysts in fecal smears. Another advantage was that oocysts could readily be detected in fecal samples stored as long as 120 days in dichromate solution. Using the dichromate solution floatation technique, it was shown that shedding of cryptosporidial oocysts in feces occurred from two to 20 days after oral challenge but was not always accompanied by diarrhea. Infection with cryptosporidia was confirmed in two calves at necropsy. Calves which were not sacrificed for portmortem examination recovered without treatment. The dichromate solution floatation technique is simple, rapid, inexpensive and should facilitate detection of cryptosporidia by diagnostic laboratories and some veterinary practitioners.

The yield of colostrum and colostral gammaglobulins in beef cows and the absorption of colostral gammaglobulins by beef calves.

The details of a small study on the volume of colostrum and colostral gammaglobulins produced by beef cows at the first milking immediately after calving and the serum concentrations of passively acquired colostral gammaglobulins attained by beef calves are reported. The mean volume of colostrum and colostral gammaglobulins produced were 2990 +/- 2100 (SD) mL and 314.75 +/- 211.8 (SD) g respectively. The mean serum concentrations of passively acquired colostral gammaglobulins were 29.9 +/- 10.1 (SD) ZST units (n = 40, Ranch A) and 19.2 +/- 7.5 (SD) ZST units (n = 42, Ranch B). The main reason for this difference appeared to be nutritional; the cows on ranch B being in much poorer condition and almost certainly producing less colostrum than those on ranch A.

The influence of feedlot management on an outbreak of bovine respiratory disease.

Epidemic curves, odds ratios and chisquare were used to investigate an epidemic of respiratory disease in a pen of feedlot cattle. The cattle were divided into four groups by an eartag number received at processing. Data for each group were collected from feedlot records describing purchase, transportation and processing histories, daily feeding methods, daily pen movements, daily diagnoses, treatment and mortality rates. These data were used to describe the effects of market origin and feeding management on the levels and distribution of respiratory disease.The three groups of cattle purchased from auction markets and started on high levels of grain in their rations were determined to be 6.3 times (P<0.0005) more likely to be treated for any disease, 4.9 times (P<0.0005) more likely to be treated for respiratory disease, 12.7 times (P<0.0025) more likely to die, and 6.7 times (P<0.0471) more likely to die with respiratory disease than the group made up primarily of farm-assembled heifers and started on a 10% grain ration with time for adjustment to grain.

Therapeutic and prophylactic effects of some antibiotics on experimental pneumonic pasteurellosis.

Pneumonic pasteurellosis was produced in cattle seronegative for bovine herpes virus-1 and Pasteurella haemolytica using their respective aerosols four days apart. When treated with four daily intravenous oxytetracycline injections one day prior, same day as and 24 hours after P. haemolytica aerosols cattle experienced a reduced mortality. Prophylactic sustained action antibiotics given 24 hours prior to the P. haemolytica aerosol also reduced mortality, however there appeared to be a variation associated with the products used.

Enterotoxigenic Escherichia coli infections in newborn calves: a review.

Diarrhea caused by enterotoxigenic Escherichia coli is an infectious bacterial disease of calves that occurs during the first few days of life. The Escherichia coli that cause the disease possess special attributes of virulence that allow them to colonize the small intestine and produce an enterotoxin that causes hypersecretion of fluid into the intestinal lumen. These enterotoxigenic Escherichia coli are shed into the environment by infected animals in the herd and are ingested by newborn calves soon after birth. There is some natural immunity to enterotoxigenic Escherichia coli; however, it often fails to protect calves born and raised under modern husbandry conditions. Hence, methods have been developed to stimulate protective immunity by vaccination of the dam. The protective antibodies are transferred passively to calves through the colostrum.

Prevalence of rotavirus and coronavirus antigens in the feces of normal cows.

The prevalence of rotavirus and coronavirus shedding by adult cows was investigated using capture enzyme-linked immunosorbent assays. Fecal samples from 121 cows in a single herd were tested for the presence of rotavirus and coronavirus, either free or complexed with immunoglobulin. Free rotavirus was not detected in any samples while rotavirus-immunoglobulin complexes were detected in 53 of 121 (44%) samples tested. In contrast, free coronavirus was detected in six (5%) samples and coronavirus-immunoglobulin complexes were detected in 85 (70%) of the samples tested. Thus it appears that subclinical infection of cows by either of these viruses is common, possibly providing a source for infection of the neonate. These assays may therefore provide important information regarding the epidemiology of enteric virus infections and suggest means of improving management to prevent epidemics of neonatal diarrhea.

A comparison of histopathological changes in calves associated with K99- and K99+ strains of enterotoxigenic Escherichia coli.

Enterotoxigenic colibacillosis was experimentally produced in four colostrum-deprived calves given 10(10) Escherichia coli strain 210 (serotype 09+:K30+:K99-:F41-:H-) orally and the histopathological changes compared to those seen in colostrum-fed calves infected in an earlier study with strain B44 (serotype 09+:K30+:K99+:F41+:H-). Escherichia coli strain 210 caused diarrhea, atrophic villi with cuboidal epithelium, and focal accumulations of a few neutrophils in the dome villi above Peyer's patches but neither the clinical nor the histopathological changes were as pronounced as with strain B44. The extent and distribution of adherence to the mucosal surface differed between the two strains. Strain B44 adhered as a continuous layer over most of the absorptive epithelial surface of both the jejunum and ileum. Adherence of strain 210 was restricted to the ileum and the bacteria often adhered focally in "clumps" rather than as a continuous layer, especially on the distal half of the villous surface.

The control of acute undifferentiated diarrhea of newborn beef calves.

A management system designed to control diarrhea in newborn beef calves has been outlined. The calving herd is moved from the wintering grounds to previously prepared, comfortable calving grounds. The herd is divided into small subunits for surveillance of calving. The calving areas should be well-bedded, and a natural or artificial windbreak should be provided. All parts of the calving grounds should be readily accessible. Every effort is made to ensure that calves receive a liberal supply of colostrum within hours after birth. Obstetrical assistance is provided as necessary to ensure that all calves are born vigorous. At 24 hours of age, the cow-calf pair is moved out of the calving grounds to the nursery pasture. Diarrheic calves are moved to an isolation area and treated accordingly. The system is successful because the newborn calf is moved from contaminated areas of high population density to areas of lower population density that are less contaminated with the common enteropathogens. Under certain circumstances, the vaccination of the dam in late pregnancy with E. coli K99+ bacterins may be beneficial as an aid to the control of the disease.

Monoclonal antibody passive hemagglutination and capture enzyme-linked immunosorbent assays for direct detection and quantitation of F41 and K99 fimbrial antigens in enterotoxigenic Escherichia coli.

Production of diarrhea in neonatal calves by enterotoxigenic Escherichia coli depends on its ability to attach to the epithelial cells of the intestine via surface adhesins called pili or fimbriae and to secrete enterotoxins. The most important of these fimbriae are designated K99 and F41. We produced and characterized a murine monoclonal antibody specific to F41. This monoclonal antibody and a K99-specific monoclonal antibody were used to develop sensitive and specific passive hemagglutination and capture enzyme-linked immunosorbent assays (ELISAs) for detection and quantitation of F41 and K99 antigens in E. coli cultures and culture supernatants. The capture ELISA systems exhibited excellent sensitivity and specificity, whereas the passive hemagglutination systems appeared to be oversensitive. The ability of the capture ELISAs to detect K99 and F41 fimbrial antigens in fecal specimens from calves was evaluated. Fimbrial antigens were detected in six of six specimens from scouring calves but not in four of four specimens from nonscouring calves.