Normal Adult Cattle Research Articles

Osteofluorosis in free ranging California sea lions (Zalophus californianus)

We report the first case series of osteofluorosis in free ranging marine mammals and documentation of baseline bone fluoride concentrations in this population; however, the exact source is not known. A stranded subadult male California sea lion, one subadult male carcass, and one partial carcass, presumably adult male, demonstrated osseous (bony) proliferations on various appendicular and axial bones. The most severe lesions were from the index case with symmetrical segmental circumferential periosteal new bone proliferation on the diaphysis of both femurs and humeri. This case, and the second case, also had significant segmental periosteal new bone proliferation on the mandibles as well as increased tooth wear with dull enamel. Total fluoride concentrations in various bones of the cases ranged from 3,700 to 9,700 ppm dw, with a mean of 5,431 ppm dw, approximately twice the concentration in adult cattle with osteofluorosis. Ribs from four control California sea lions of various ages and both sexes had an average total fluoride of 1,575 ppm dw, which is three times as high as the bone fluoride concentration of normal adult cattle, but four times lower than the mean rib fluoride concentrations in the cases. This is the first report of osteofluorosis in free ranging marine mammals and documentation of baseline bone fluoride concentrations in this population. The exact source of the excessive fluoride levels is not known as male California sea lions have a large range and varied diet.

Laboratory findings of tracheal wash and bronchoalveolar lavage in normal adult dairy cattle

ABSTRACT Tracheal Wash (TW) and bronchoalveolar lavage (BAL) are not well documented in adult cattle and published studies were undertaken on diseased animals aiming at isolating the causative agent of respiratory diseases. This study was undertaken to establish the normal cytological and microbiological findings of TW and BAL in normal adult cattle and compare between the laboratory analysis findings of both techniques. Eighteen healthy adult cattle were divided into two groups. BAL was completed the first group (n = 10) and TW was completed the second group (n = 8). Normal cytological findings of TW in healthy adult cattle were found to be primarily neutrophils (42.7 ± 35.7)% intermixed with lower numbers of columnar epithelial cells (26.2 ± 34.8)%, alveolar macrophages (14.7 ± 15.4)%, squamous epithelial cells (10 ± 28.6)%, lymphocytes (6.8 ± 16.1)% and rarely seen eosinophils (0.1 ± 0.0)% with a total cell count of 457 ± 310.1 cells/µl. Cytological findings of BAL were found to be mainly alveolar macrophages (94.3 ± 3.9)%, intermixed with lower number of neutrophils (4.3 ± 3.9)% and lymphocytes (1.3 ± 1.2)% and rarely seen eosinophils (0 ± 0.1)% with a total cell count of 238 ± 169 cells/µl. There are remarkable differences in TW and BAL cytological findings and the degree of contamination was higher in TW samples than that seen in BAL lavage.

A comparison of titers of anti-Brucella antibodies of naturally infected and healthy vaccinated cattle by standard tube agglutination test, microtiter plate agglutination test, indirect hemagglutination assay, and indirect enzyme-linked immunosorbent assay.

Aim:We determined the antibody response in cattle naturally infected with brucellosis and normal healthy adult cattle vaccinated during calf hood with strain 19.Materials and Methods:The antibody titers were measured by standard tube agglutination test (STAT), microtiter plate agglutination test (MAT), indirect hemagglutination assay (IHA), and indirect enzyme-linked immunosorbent assay (iELISA) as per standard protocols.Results:The mean STAT titers were 1.963±0.345 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was extremely significant (p<0.0001). The mean MAT titers were 2.244±0.727 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was very significant (p<0.005). The mean IHA titers in infected cattle were 2.284±0.574, and those in healthy vaccinated cattle were 1.200±0.155. The difference was extremely significant (p=0.0002). However, the difference in mean iELISA titers of infected cattle (1.3678±0.014) and healthy vaccinated cattle (1.367±0.014) was non-significant. The infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals. However, it cannot be ascertained whether these antibodies are due to vaccine or response to infection. Since the infected animals had been vaccinated earlier, the current infection may suggest that vaccination was unable to induce protective levels of antibody. The heightened antibody response after infection may also indicate a secondary immune response to the antigens common to the vaccine strain and wild Brucella organisms.Conclusion:The brucellosis infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals.

Development and application of one-step multiplex reverse transcription PCR for simultaneous detection of five diarrheal viruses in adult cattle

A one-step multiplex reverse transcription (RT)-PCR method was developed for the simultaneous detection of five viruses causing diarrhea in adult cattle: bovine group A rotavirus (GAR), bovine group B rotavirus (GBR), bovine group C rotavirus (GCR), bovine coronavirus (BCV), and bovine torovirus (BToV). The detection limit of the one-step multiplex RT-PCR for GAR, GCR, BCV, and BToV was 102, 100, 101, and 102 TCID50/ml, respectively, and that for GBR was 106 copies/ml. The one-step multiplex RT-PCR with newly designed primers to detect GAR had higher sensitivity than a single RT-PCR with conventional primers, with no false-positive reactions observed for ten other kinds of bovine RNA viruses To assess its field applicability, 59 of 60 fecal samples containing one of these five viruses from all 25 epidemic diarrhea outbreaks in adult cattle were positive in the one-step multiplex RT-PCR assay. Furthermore, using four additional fecal samples containing two viruses (GBR and BCV or BToV), two amplified products of the expected sizes were obtained simultaneously. In contrast, all 80 fecal samples lacking the five target viruses from normal adult cattle were negative in the multiplex assay. Taken together, our results indicate that the one-step multiplex RT-PCR developed here for the detection of GAR, GBR, GCR, BCV, and BToV can be expected to be a useful tool for the rapid and cost-effective diagnosis and surveillance of viral diarrhea in adult cattle.

Evaluation of the Perkins handheld applanation tonometer in horses and cattle

The objective of this study was to evaluate and validate the accuracy of the Perkins handheld applanation tonometer for measuring intraocular pressure (IOP) in horses and cattle. Both eyes of 10 adult horses and cattle were evaluated in a postmortem study. The eyes from 10 clinically normal adult horses and cattle were also examined after bilateral auriculopalpebral nerve block and topical anesthesia for an in vivo study. IOP was measured postmortem using direct manometry (measured with an aneroid manometer) and tonometry (measured with a Perkins handheld applanation tonometer). The correlation coefficients (r2) for the data from the postmortem manometry and Perkins tonometer study were 0.866 for horses and 0.864 for cattle. In the in vivo study, IOP in horses was 25.1 ± 2.9 mmHg (range 19.0~30.0 mmHg) as measured by manometry and 23.4 ± 3.2 mmHg (range 18.6~28.4 mmHg) according to tonometry. In cattle, IOP was found to be 19.7 ± 1.2 mmHg (range 18.0~22.0 mmHg) by manometry and 18.8 ± 1.7 mmHg (range 15.9~20.8 mmHg) by tonometry. There was a strong correlation between the IOP values obtained by direct ocular manometry and the tonometer in both horses and cattle. Our results demonstrate that the Perkins handheld tonometer could be an additional tool for accurately measuring IOP in equine and bovine eyes.

Comparison and verification of BSE surveillance in USA and Japan

The objectives of this study are to (1) compare the BSE surveillance systems of Japan and the United States (US), and to (2) validate the US enhanced BSE Surveillance program. This study compares the BSE surveillance systems in Japan and the US, specifically focusing on the procedures of initial test, diagnosis and confirmation. The study further examines the validity of statistical conclusions made in the US enhanced surveillance program based on the data collected from the BSE inspection performed by the Ministry of Health, Labor and Welfare of Japan (MHLW) and the Ministry of Agriculture, Forestry and Fisheries of Japan (MAFF) between October 18,2001 and July 31, 2004. The inspection targeted all slaughtered healthy and high risk cattle. The US enhanced surveillance program assumes no BSE occurrence in the normal adult cattle population and thus its inspection focuses only on high risk cattle. The BSE inspection performed in Japan, however, revealed that 0.00022% of the normal adult cattle were BSE-infected using the US criteria. Assuming that the same ratio of cattle was BSE-infected in the US, the Japan finding indicates that approximately 30% of the US slaughtered normal cattle population aged 30 months and over needs to be tested to satisfy the statistical condition used by the US (i.e., 99% confidence level). On the other hand, in order for Japan to perform the surveillance with a 99% confidence level (the statistical condition used by US), Japan needs inspect: (1) 60,539 high risk cattle (i.e., 60% of 100,583 high risk cattle); (2) 78% of normal adult cattle aged 30 months and over (1,088,589/1,387,522) and (3) 90% of normal cattle aged 30 months or less (1,845,138/2,050,154). The US enhanced surveillance program launched in July 2004 is based on the premise that no BSE occurs in normal adult cattle population. In Japan, however, BSE cases satisfying the US criteria have been found among the normal adult cattle. This fact suggests that the US needs to consider inspections targeting the normal adult cattle. This fact suggests that the US needs to consider inspections targeting the normal adult cattle in the future. At the same time, for more efficient surveillance, Japan may need to consider BSE inspections targeting the high risk cattle population with a higher confidence level and normal adult cattle with a lower confidence level.

Evaluation of a commercial ELISA for detecting serum antibody to Neospora caninum in cattle

A commercially available serum antibody detection enzyme-linked immunosorbent assay for Neospora caninum in cattle was evaluated against an immunofluorescent antibody test (IFAT) by applying it to 397 sera from normal...

Distribution of T-lymphocyte Subpopulation in Blood and Spleen of Normal Cattle and Cattle with Enzootic Bovine Leukosis

Immunocytochemical and flow cytometry techniques were used to examine T-lymphocyte subpopulations in peripheral blood lymphocytes (PBLs) and spleen from cases of enzootic bovine leukosis (EBL) in adult cattle, and from normal cattle (adult and young), with a panel of monoclonal antibodies against bovine leucocyte differentiation molecules. Both in PBLs and spleen, the percentages of T-lymphocyte subpopulations (CD3+, CD4+, CD8+, and WC1+γδT lymphocytes) of EBL-affected and normal adult cattle were significantly lower than those of normal young cattle. The percentages of these T-lymphocyte subpopulations in the PBLs of adult cattle with EBL were lower than those of normal adult cattle, but the converse was true in the spleen. It is suggested that tumour immunity occurred in the spleen. Histological examination revealed no follicular hyperplasia in the spleen, and the proliferation of neoplastic cells began in the red pulp. It is concluded that the spleen is not the organ initially responsible for the transformation of EBL lymphoma and that neoplastic cells migrating from peripheral blood are metastatic.

Comparative analyses of peritoneal fluid from calves and adult cattle

SUMMARY Reference values for hematologic variables change with increasing age in cattle. Therefore, the purpose of the study reported here was to describe the peritoneal fluid constitutents of clinically normal young calves, and to compare cellular concentration and distribution in blood and peritoneal fluid of young calves with those of adult cattle. Eight healthy 8-week-old male Holstein calves and 8 healthy 3- to 8-year-old Holstein cows were studied. Peritoneal fluid was collected from calves along the ventral midline, 4-cm cranial to the umbilicus. Abdominocentesis was performed in the region of the lower right flank in adult cattle. Correlation analysis, using the Pearson's correlation coefficient, and regression analysis were performed for blood and peritoneal fluid data from calves. Data from calves were compared with those of cows, using Wilcoxon's rank sum test. A P value &lt; 0.05 was considered significant for all tests. Calves had significantly lower blood eosinophil count (P &lt; 0.003) and plasma protein concentration (P &lt; 0.001) than did cows. Calves had significantly higher peritoneal fluid nucleated cell (P &lt; 0.05) and mononuclear cell (P &lt; 0.05) counts, but lower peritoneal fluid eosinophil cell count (P &lt; 0.003) than did cows. For calves, nucleated cell and lyphocyte cell counts in the blood had a high, positive correlation with those of peritoneal fluid. However, the prediction equation for nucleated cell count accounted for a modest proportion of variability. A prediction equation for peritoneal fluid lymphocyte cell count was established. On the basis of results of this study, reference ranges established for peritoneal fluid constituents of clinically normal adult cattle may not be appropriate for interpretation of peritoneal fluid analysis of calves.

Composition and analysis of cerebrospinal fluid in clinically normal adult cattle

Summary Cerebrospinal fluid and serum were obtained from 16 clinically normal adult cows (11 dairy, 5 beef). Sodium, potassium, magnesium, total protein, and albumin concentrations, osmolality, and lactate dehydrogenase and creatine kinase activities, were quantified in csf and serum. Total and differential cell counting, protein electrophoresis, and IgG quantification were performed on csf. Statistical analyses of these variables, including mean, sem, range, and 95% confidence intervals, were performed. Effects of blood contamination were evaluated, and were found to be negligible for all measured constituents. Correction factors for csf creatine kinase and lactate dehydrogenase activities accounting for cellular contamination were developed. Total nucleated cell count was similar to counts in csf of other species, but higher than values in healthy people. Differential leukocyte count in csf was similar to that reported in csf of other domestic animals: mostly lymphocytes, fewer monocytoid cells, and scant neutrophils. Cerebrospinal fluid protein concentration was higher than concentration reported for dogs, goats, and people, but was similar to values reported for horses. Beef cows had higher csf total protein concentration than did dairy cows; also, beef cows had higher csf γ-globulin concentration. The concentration of sodium in csf was slightly higher than the value in serum, and potassium concentration was lower than the value in serum. In contrast to studies of human beings, csf osmolality was generally less than serum osmolality in the cows studied. Reference values for csf electrolyte concentrations and osmolality are useful for diagnosis of salt poisoning and for assessment of the effects of fluid therapy. Magnesium concentration was lower in csf, compared with serum. Reference values may be useful for diagnosis of grass tetany. Glucose concentration in csf was variable, compared with serum concentration; sometimes, it was similar, lower, or even higher than serum glucose concentration. This variation reflects a more complete equilibration of glucose between csf and blood, owing to the lower and more stable blood glucose concentration in cows. Creatine kinase activity in csf was markedly less than, and was not correlated with, serum creatine kinase activity. Cerebrospinal fluid lactate dehydrogenase activity was markedly lower than serum lactate dehydrogenase activity. Compared with lactate dehydrogenase, creatine kinase activity had a wider range in these healthy cows; therefore, the former enzyme has higher specificity and sensitivity for diagnosis of diseases affecting the cns.

Lymphocyte blastogenesis and cellular cytotoxicity in a congenital infection of bovine fetuses related to epizootic bovine abortion

The disease referred to as epizootic bovine abortion (EBA) was experimentally induced in bovine fetuses. Dark-field microscopy was used to detect congenital infection with an unclassified spirochaete-like organism. Some of the fetuses collected at abattoirs were also found to be naturally infected with a morphologically similar microorganism. Blood counts and organ weights were correlated with the presence of the microorganism. Lymphocyte blastogenesis increased, the result of in vivo stimulation among the infected fetuses. Phytomitogens (phytohaemagglutinin, concanavalin A and lipopolysaccharide) also stimulated greater responses in infected fetuses when compared to results in normal fetuses. Cellular cytotoxicity was examined by the single cell assay and results indicated that there were fewer cytotoxic lymphocytes among the diseased fetuses. The infected abattoir-collected specimens were obtained from clinically normal adult cattle, and the immunological changes in these fetuses were closely characterised with those of the EBA diseased fetuses. These naturally infected fetuses showed signs of a mild infectious disease.

Ontogeny of mitogen responsive lymphocytes in the bovine fetus

Bovine fetal lymphocytes were examined for their ability to respond to the mitogens phytohemagglutinin (PHA), Concanavalin A (ConA) and pokeweed mitogen (PWM) in the lymphocyte blastogenesis test (LBT). PHA-, Con A- and PWM-responsive lymphocytes appeared simultaneously at 75–80 days of gestation. The response increased with age of the fetus until, by 120 days of gestation, the response to PHA, Con A and PWM of many fetuses was in the range of values obtained with lymphocytes from normal adult cattle.

牛の血中シアル酸およびムコ蛋白の測定法と臨床応用

Sialic acid (SA) in bovine blood was assayed by five methods: the enzyme, the resorcinol, the thiobarbituric acid, Ehrlich's, and Hess'. There were highly significant statistical correlations among the values obtained by these methods. The plasma concentration of SA measured by the enzyme method and that of mucoprotein (MP) measured by the Coomasie Brilliant Blue G-250 method were 51.6±7.6mg/dl and 111.3±33.4mg/dl, respectively, in 136 healthy adult cattle. The plasma level of SA was higher in 138 bovine neonates during 14 postnatal days than in normal adult cattle, and that of MP in the former during 35 postnatal days.The serum concentrations of SA and MP were measured in 206 cattle affected with some inflammatory diseases diagnosed by meat-inspecting or practicing veterinarians. They were much higher in cattle with pyemia and endocarditis verrucosa and higher in those with liver abscess, nephritis, arthritis and mastitis than in normal cattle. They showed a difinite pattern in cattle having received laparotomy during a postoperative period till repair. As indicators for the diagnosis of an inflammatory disease and for the condition of postoperative course in cattle, the assay of both substances seemed to be very useful.

Serum pepsinogen levels andOstertagia ostertagipopulations in clinically normal adult beef cattle

Serum pepsinogen levels and Ostertagia ostertagi populations in clinically normal grass-fed bullocks were investigated in three groups of 10 prime cattle aged between 2.5 and 2.75 years slaughtered in late summer (February), early autumn (March) and late autumn (May) respectively. Apart from occasional foci of mucosal hyperplasia abomasa were grossly normal. Serum pepsinogen levels ranged between 0.2 and 2.5 i.u./l with group means of 1.4, l.5 and 1.3 i.u./l. O. ostertagi counts ranged between 0 and 5,194 with group means of 734,630 and 701 worms. The composition of the worm populations varied with a higher proportion of adults recovered in February and very few worms from most cattle in March, suggesting the termination of a parasite generation. An increase in numbers of early fourth-stage larvae in May indicated exposure to a new generation. These changes were not reflected in the pepsinogen levels. The findings are discussed in relation to the adequacy of the pepsinogen assay as a diagnostic aid in field infections, animal age, and correlations between pepsinogen levels and parasite populations.

Colostral transfer of gamma glutamyl transpeptidase in calves

Serum gamma glutamyl transpeptidase (GGT) levels in blood samples taken from normal calves which bad suckled colostrum were much higher than those found in healthy adult cattle. Levels of over 60 times the normal adult level were observed. These high levels of GGT took approximately 5 weeks to decline to adult values. Calves which appeared to have not received or absorbed colostrum had GGT levels which would be considered normal in adult cattle. A calf with serum gamma globulin levels which indicated an intermediate amount of colostrum absorption had a level of GGT which was intermediate between that expected for normal adult cattle and that found in calves which had more fully absorbed colostrum. The mean GGT level observed in colostrum from 6 newly-calved cows was over 800 times the mean serum GGT level of the same 6 cows. It therefore appears most likely that GGT is concurrently absorbed with colostrum by calves and this gives rise to the very high levels seen in normal calves. Calves with very high levels of serum GGT also had raised levels of alkaline phosphatase (AP) which slowly declined but never reached normal adult levels within the 53-day period of observation. It therefore appears that high levels of both GGT and AP are achieved by calves at the time of colostrum absorption and it is concluded that clinical interpretation of serum GGT and AP levels in young calves is closely dependent upon parallel knowledge of their serum gamma globulin levels.

Agglutination of Brucella abortus cells by sera from cattle experimentally infected with Escherichia coli

Infection of normal adult cattle and sporadic serological Brucella abortus reactors, which no longer had diagnostic titers to B. abortus, with a culture of Escherichia coli isolated from a cow (or its environment) resulted in the production of a primary IgM serum antibody response to both B. abortus and E. coli. Cattle injected with heat killed E. coli and guinea pigs or young calves lacking natural agglutinins to B. abortus injected with live E. coli, produced serum antibody only to E. coli. The subsequent reinjections of live E. coli into the former two groups of cattle resulted in all but one animal in each group producing IgM ‘secondary’ responses to B. abortus of decreasing magnitude, while the anti- E. coli responses increased and eventually switched to synthesis of IgG class antibody. The remaining two animals produced a series of ‘secondary’ responses of IgM antibody to B. abortus similar in amplitude and duration to the primary immune response. The anti- E. coli agglutinins of these two animals increased in titer and IgG class antibody to E. coli was evident after several injections of E. coli. These results indicate that infection of cattle by E. coli can cause a problem in the serological fiagnosis of B. abortus infection. Speculations on the cause of this cross-reaction and ways of minimizing misdiagnosis are discussed.

Serological relation between calf diarrhea coronavirus and hemagglutinating encephalomyelitis virus.

SummaryNeutralizing (NT) and hemagglutination-inhibiting (HI) antibodies to calf diarrhea coronavirus (CDCV) and hemagglutinating encephalomyelitis virus of swine (HEV) (strain 67 N) were detected in high proportions of normal adult cattle and pigs in Japan. Since comparison of NT and HI titers in the serum samples suggested an antigenic difference between the viruses, cross NT and HI tests of these viruses were carried out with antisera raised in rabbits. The homologous NT titers were markedly higher than the heterologous titers. In HI tests essentially the same results were obtained. These findings indicate the presence of a marked difference in antigenic make-up between CDCV and HEV. NT and HI tests can clearly differentiate the viruses, although there is some cross reaction.

The significance of elevated plasma creatine phosphokinase activity in muscle disease of cattle

Creatine phosphokinase activity was measured in the plasma of normal adult and young cattle, in heifers and their calves during the period after parturition, in cattle after liver or muscle biopsy and in calves after an intramuscular injection of Terramycin. The same enzyme was measured in cattle with experimentally produced myodegeneration. The half-life of creatine phosphokinase in the plasma of cattle was compared with that of aspartate aminotransferase and lactate dehydrogenase. The value of this enzyme as an indicator of muscle disease is discussed.

Further studies on the antigenic properties and distribution of the putative bovine leukemia virus.

The C-type viruses found in long-term cultures. New Bolton Center (NBC) cell lines, of peripheral lymphocytes from leukemic cattle and in short-term cultures of bovine buffy coat(BC) cells share an immunofluorescent(IF)antigen detected in the cytoplasm of infected cells as well as an antigen demonstrable in gel diffusion experiments. Therefore the viruses from these cultures most likely represent different isolates of the putative bovine leukemia virus (BLV). The BLV precipitin antigen is analogous to the group specific (gs) antigens of the leukemia viruses of other species in that it is soluble, ether resistant, and apparently located within the virion. These observations, together with results showing that the specificity of the BLV precipitin antigen differs from that of the gs antigen of other mammalian leukemia viruses, indicate that the former antigen represents the intraspecies (gs-1) determinant of BLV. Antibodies to the precipitin viral antigen were found in 82% of cattle with leukemia and in 40% of clinically normal adult cattle in multiple-case herds. These groups of animals also had fluorescent antibodies to the virus, but with significantly higher frequencies (100% and 76%, respectively). On the other hand, in leukemia-free herds, precipitating antibodies were not found and the incidence of fluorescent antibodies was only 3%.

The Effect of Breed and Age on the Distribution of Conglutinin and Immunoconglutinin in Normal Cattle

Summary A total of 300 normal cattle of different breeds and age were assayed Jor serum conglutinin (K) and immunoconglutinin (IK) oner a period of 6 months, covering a cross section of wet and dry seasons. Adult Zehn cattle showed the highest levels of K. Adult Hereford cattle had widely scattered values of K and some (6.5%) appeared in the low-litre range. None of the crossbred Hereford-Zebu adult cattle were detected in the low K level range. Calves attained normal adult K levels by 6 months of age regardless of their breed. The K levels of normal adult cattle ranged between o and 2500 100% conglutinating units (K100/ml). One of the Junctions of K may be the regulation of complement levels. IK was detected in only 1.5% of all the samples tested and only at low titre. This protein was restricted to adult cattle. It is suggested that the occasional appearance of IK in normal populations is a reflection of antigenic experience.