Mutation scan screening of Echinococcus granulosus isolates of Indian origin

During the present investigation a total of forty Indian animal isolates were screened by single strand conformation polymorphism (SSCP) collected from sheep, goat, cattle and buffalo. The result of the study indicated that nuclear variants of Echinococcus granulosus were present in both small and large ruminants. SSCP phenotypes of AgB, intron of actin II and Hbx-2 have been deduced. Presence of nuclear variants due to mutation of E. granulosus has been discussed depending on hypotheses imparted earlier in literature. High polymophism of AgB demands further investigation because the gene is related with immune evasion and infectivity. This communication reports for the first time the comparative profile of Indian goat, sheep, cattle and buffalo isolates of E. granulosus complex.     

Differentiation antigens on bovine mononuclear phagocytes identified by monoclonal antibodies

Five monoclonal antibodies (MAb) produced against cell surface antigens on bovine mononuclear phagocytes (MPh) were characterized. None of the MAb recognized erythrocytes, thrombocytes, B lymphocytes or resting or activated T lymphocytes. Two MAb (IL-A22 and IL-A24) reacted with the majority of monocytes and granulocytes in peripheral blood, with 20-40% bone marrow cells comprising myelo-monocytic cells, and with a proportion of mature macrophages. Reactivity of the remaining three MAb was restricted to MPh: one of these (IL-A25) was apparently specific for pulmonary macrophages, whereas the molecules recognized by the other two (IL-A23 and CH16A) were expressed on subpopulations of blood monocytes and tissue macrophages. None of the MAb inhibited adherence of MPh to plasma-coated gelating surfaces or Fc-mediated rosette formation. One of the MAb, IL-A24, which reacts with MPh and granulocytes, inhibited antigen-specific proliferative response or peripheral blood mononuclear leukocytes (PBM) to the soluble antigen, keyhole limpet hemocyanin (KLH) but did not inhibit responses to concanavalin A or allogeneic leukocytes. This MAb was shown to react with two polypeptides of approximately 75 kD and 110 kD on the surface of peripheral blood monocytes.     

Experimental bovine group-B streptococcal mastitis induced by strains of human and bovine origin

Intramammary infections with Group-B streptococci of human and bovine origin were produced experimentally in cows. The initial cytological response was more rapid to the human than to the bovine strain (Table I), while at later stages the pathological changes induced by the two infections were much the same (Fig. 1). The initial clinical reaction was more acute to the "human" than to the "bovine" infections and the average daily loss of milk was greater in cases of "human" infection than in cases of "bovine" infection (Table II). In contrast to the "bovine" infections the "human" infections showed a pronounced tendency to spontaneous clearance. The rate of excretion of Group-B streptococci with the milk was lower for the "human" than for the "bovine" infections (Table III). The special mode of reaction of the bovine udder against infections with Group-B streptococci of human origin may, in part, explain why such infections have a lower tendency to spread within a herd than infections with bovine strains of B-streptococci.     

Separation by carrier-free electrophoresis of subpopulations of bovine alveolar macrophages

Carrier-free electrophoresis was used to separate subpopulations of bovine alveolar macrophages according to their electrical surface charges. The electrophoretic mobility of the macrophages changed significantly as a function of the time of cultivation in vitro. After 20 h, three subgroups were identified which differed in their specific contents of lysosomal hydrolases, alpha-naphthyl acetate esterase, and alkaline phosphodiesterase. By comparing the findings with results from the literature it was concluded that the subgroups correspond to different stages of maturation and that the fraction moving fastest may be the most mature fraction. The results were not influenced by phagocytosis and the presence or absence of Ca2+.     

Variations in infestations shown by dogs following a fixedfeeding of hydatid scolices (Echinococcus granulosus)

Sir,—In the course of a survey of apparently healthy bobby calves at the Auckland Farmers Freezing Company works at Horotiu, Hamilton, an opportunity was taken to examine certain groups of calves during the months of August and September, 1964. These calves were (a) dead on arrival, (b) condemned for immaturity as judged by the weight (<45lb), moist appearance of the navel and lack of wear of the foot pads, or (c) condemned as being unfit — i.e., showing some obvious deviation from normal clinical appearance, such as collapse, stupor, diarrhoea or suffering from trauma.

     

Diagnosis of bovine brucellosis by enzyme immunoassay of milk

Enzyme-immunoassays using lipopolysaccharide as antigen were developed for the detection of bovine IgG1, IgG2 or IgA Brucella antibodies (Ab) in milk. The results of these tests were compared with those of the milk ring test (MRT) by analyzing 3212 bulk milk samples from farms located in regions where brucellosis is prevalent. Among the 105 herds detected by ELISA and/or MRT, 29 infected herds were detected by ELISA only. The 40 MRT-positive herds were also ELISA positive. Five herds became infected during the study and were detected by ELISA 15 days to 6 months prior to detection by MRT. The ELISA IgG1 titration (IgG1 ELISA) detected 92.8% of the herds found positive in the three ELISA assays. The concomitant use of IgA ELISA raised the sensitivity to 100% but slightly decreased the specificity. The IgG2 ELISA did not improve the diagnosis. The sensitivity of MRT and IgG1 ELISA was compared by testing successive dilutions in negative milk of 110 individual MRT positive milks samples. On average, IgG1 ELISA was 22 times more sensitive than MRT.     

Setaria digitata in cattle of Thailand identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis.

Adult Thai Setaria worms collected from cattle which were bred, housed and slaughtered in Thailand were morphologically identified as Setaria digitata. Furthermore, in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) adult Thai S. digitata had the same protein profiles as adult Japanese S. digitata, but did not possess the protein with a molecular size of 69 kDa which was confirmed in adult S. marshalli. In addition, there were no differences in the protein profiles between male and female S. digitata. In point of the distribution pattern of the proteins ranging from 73 to 64 kDa revealed by 2D-PAGE, there were no differences between Thai and Japanese S. digitata, and between male and female worms of the species.     

Identification of envelope and nucleocapsid proteins of infectious bovine rhinotracheitis virus by SDS-polyacrylamide gel electrophoresis

Infectious bovine rhinotracheitis (IBR) virus was purified by rate zonal and isopycnic centrifugation in potassium tartrate gradients. Viral nucleocapsids were isolated from purified virions by treatment with the nonionic detergent Triton X-100 followed by high speed centrifugation. This treatment was shown to produce a suspension of 74% completely de-enveloped nucleocapsids, 24% incompletely de-enveloped nucleocapsids, and 2% whole virions. The viral nucleocapsids contained DNA and banded at a density of 1.25 g/cm3. Analysis of the viral polypeptides by gradient SDS-polyacrylamide gel electrophoresis revealed that 33 virion proteins, ranging in molecular weight from 13,000 to 275,000 dalton, were present in the complete virus particle. Detergent treatment of the virus quantitatively removed two of the major proteins (vp8, 90,000 dalton, and vp13, 73,000 dalton) and partially removed eleven other proteins. Fifteen viral polypeptides appeared to remain firmly associated with the viral nucleocapsids.     

Hepatic enzyme changes in bovine hepatogenous photosensitivity caused by water-damaged alfalfa hay

In the winter of 1983, practitioners reported extensive photosensitization in 7 herds of cattle. All herds had a history of having been fed water-damaged alfalfa hay. A cow from one herd was referred to the veterinary teaching hospital at Oklahoma State University. In this herd of approximately 40 adult Polled Herefords, all cattle had had some degree of clinical involvement over the past 4 to 6 weeks. Clinical signs included scaling and erythema of sparsely haired skin, muzzle, and teats, as well as icterus, anorexia, and weight loss. One cow died, and the remaining cattle recovered over an 8- to 10-week period after removal of the hay from the ration. In the referred cow, values for total and conjugated bilirubin, BUN, creatinine, sorbitol dehydrogenase, serum alkaline phosphatase, serum aspartate transaminase, and serum gamma-glutamyl transferase were higher than normal. In the herd of origin, extremely high serum gamma-glutamyl transferase values (180 to 1,400 IU/L) persisted (normal, 2 to 35 IU/L). Feeding the same alfalfa hay to 2 clinically normal cows reproduced the syndrome. The characteristic hepatic lesion was bile duct necrosis, with secondary bile duct hyperplasia.     

Direct enzyme immunoassay of progesterone in bovine plasma

The present study was undertaken to develop a novel, practical and simple procedure for enzyme immunoassay (EIA) of plasma progesterone in cows. Diluted plasma was heated for 70°C for 30 min and applied directly to wells of a microtitre plate without extraction. Then plasma was incubated with antiprogesterone antibody and horseradish peroxidase‐labeled progesterone. The sensitivity of the assay was estimated as 4.4 pg/mL (0.11 pg/well). The intra‐assay and interassay coefficients of variation were 5.7–19.1% and 6.6–19.3%, respectively. When 0.3, 1 and 3 ng of progesterone were added to plasma, the recovery rates ranged between 79.9 and 108.4%. Only 4 h were needed to complete an assay to measure progesterone concentration. To apply the present direct EIA, progesterone concentration in plasma was assayed in crossbred cows used for the embryo transfer program. During insertion of controlled‐internal drug release (CIDR), progesterone concentrations were kept at a high level, although the removal of CIDR with treatment of dinoprost trometamine reduced progesterone concentration drastically. These results suggest that the present direct EIA is a practical and suitable method for measuring the plasma concentration of progesterone.     

Molecular typing of Staphylococcus aureus of bovine origin by polymorphisms of the coagulase gene.

Mastitis caused by Staphylococcus aureus is a disease of major economic importance to the dairy industry. Transmission occurs during milking, with chronically-infected cows acting as the major reservoirs of infection. PCR-coagulase gene typing of 151 S. aureus isolates from seven farms generated only six PCR types, with 110 (73%) isolates assigned to PCR type 1 and 23 (15.2%) isolates assigned to type 2. PCR type 1 was the predominant type on five of the seven farms, including farms in geographically separated regions of Victoria, while type 2 predominated on two farms. With the exception of the 41 isolates from one farm, all isolates were resistant to penicillin, but susceptible to other antibiotics that are routinely used to manage mastitis in dairy cattle. Nine of 11 cows with chronic S. aureus infection showed evidence of persistence of a single PCR type for periods of up to 9 months. Two different PCR types of S. aureus were isolated from the other two cows over the same period.     

Prevalence of bovine tuberculosis in slaughtered cattle identified by nested-PCR in abattoirs from two dairy areas of Ecuador

Bovine tuberculosis (bTB) is a chronic granulomatous disease that primarily affects lung tissue and lymph nodes (LN) in cattle, with economic impact on their productivity. Furthermore, it is potential zoonoses that may cause public health hazard. In this study, we evaluated the presence of bTB in two abattoirs: Cayambe and Pelileo countries located in the Ecuadorian provinces of Pichincha and Tungurahua, respectively. In total, 578 cattle were sampled (Cayambe 271 and Pelileo 307): 1,156 LN and 578 lung tissue samples were collected to apply in vitro culture and nested-PCR, respectively. The results determined a total apparent prevalence of 4.33 %, with 4.06 % at Cayambe’s abattoir and 4.56 % at Pelileo’s abattoir. Additionally, the Bayesian analysis showed a total true prevalence of 2.51 %, with 89.7 % of sensitivity and 97.6 % of specificity. The risk factors were evaluated by the use of simple logistic regressions with and without the random effect of places of origin. Associations of the origin of cattle in the selected slaughterhouses were found. The results showed an efficient method for the detection of bTB, which could identify a large number of infected animals, and the usefulness of lung tissue samples for early diagnosis of the disease was demonstrated in this study.