Characterization of infertility and bovine leukemia virus infection in beef bulls on southwestern Louisiana coastal range

One hundred and twenty-six beef bulls on southwestern Louisiana coastal range were evaluated for breeding soundness. Samples were taken to determine the incidence of bovine leukemia virus (BLV) infection, and the prepuce was cultured for potential pathogens. A high incidence (47.6%) of questionable and unsatisfactory potential breeders resulted mainly from 37.0% of the bulls exhibiting high numbers of abnormal sperm cells in the semen. Only bulls in the 4-to 5-yr age group exhibited the expected incidence of normal spermiograms. Genital campylobacteriosis was not diagnosed but there was genital trichomoniasis in three of the seven herds. Hemophilus somnus , mycoplasma and ureaplasma were isolated from the prepuce of 13.3, 48.8 and 36.7% of the bulls, respectively. Isolation of these organisms from the prepuce did not appear to be associated with abnormal spermiograms. Of the bulls studied, 34.4% had positive AGID reactions for BLV. Bulls seropositive to BLV had an increased incidence of leukocyte counts that were above the normal range. There was no apparent relationship between BLV infection and abnormal spermiograms.     

Severity-related molecular differences among nineteen strains of dengue type 2 viruses

Comparative nucleotide sequencing was carried out on dengue type 2 virus (DEN-2) strains isolated from patients in Northeast Thailand during the epidemic season in 1993. The patients exhibited different clinical manifestations ranging from dengue fever (DF) to dengue haemorrhagic fever (DHF)/dengue shock syndrome (DSS). The results classified 19 DEN-2 strains into 3 subtypes according to nonsynonymous amino acid replacements. The strain isolated from a DSS patient eliciting secondary serological response belonged to subtype I, whereas 13 strains isolated from DHF patients with secondary response and 2 strains from DF patients with primary response belonged to subtype II. On the other hand, 3 strains isolated from DF cases evoking either primary or secondary response belonged to subtype III. These results suggest that subtype III virus infection could result in clinically milder manifestation irrespective of the serological response compared with subtype I or II viruses. The RNA secondary structure predicted for the 3' noncoding region showed 4 different structures (A, B, C, and D). The result also indicates that different subtypes of DEN-2 serotypes are circulating in a single epidemic in Thailand.     

胡枝了属根瘤菌的多相分类研究

采用数值分类,全细胞可溶性蛋白电泳分析,ＤＮＡ,Ｇ＋Ｃｍｏｌ％和ＤＮＡ相关性的测定以及１６ＳｒＤＮＡＰＣＲ－ＲＦＬ分析等多相分类技术对来源于不同地区的１６种寄主的胡枝子根瘤菌进行了系统的分类研究,数值分类的结果表明,在６７％的相似性水平上,全部供试菌可以为快生型根瘤菌和慢性型根瘤菌两大群,在８０％的相似性水平上又可分为两个亚群。在此基础上,对各亚群的胡枝子根瘤菌进行了ＤＮＡ相关性的测定,以进一步证     

Frequency and distribution per species, biotypes, resistance to antibiotics and beta-lactamase production of the haemophils isolated from patients with respiratory diseases

A number of 150 samples were prelevated from respiratory tract secretions of 88 patients with respiratory infections and three healthy subjects; 162 haemophilus strains were isolated, identified and studied and the following results were obtained: H. parainfluenzae was isolated from tonsillitis and laryngitis--over 70%, bronchitis--58% and pharyngitis--56.6%; H. influenzae was isolated from pharyngitis--26.4%, bronchitis--16.1% and tonsillitis--13.6% cases; H. parahaemolyticus from bronchitis--19.3%, tonsillitis--13.6% and laryngitis. H. paraphrophilus was isolated (6.8%) from pharyngitis, tonsillitis, sinusitis, bronchitis and pulmonary abscess and H. paraphrohaemolyticus was isolated--4.5% from pharyngitis, synusitis, bronchitis and pulmonary sarcoidosis. Most of the isolates belonged to biotype II H. influenzae and biotypes II, I, III H. parainfluenzae. Haemophils were 100% sensitive to Ofloxacin and resistant to Cro--13.5%, Do--17.9%, C and Caz--22.2%, Aml--24.6%, Rd--40.7%, Amp--41.9% and Te--63.5%; varying according to the haemophilus species. H. influenzae was resistant to Do--14.2%, Caz and C--21.4%, H. parainfluenzae was resistant to Cro--11%, Do--22%, whilst H. parahaemolyticus was resistant to Do--9% and to Aml, Caz and Cro--13.6%. Haemophils isolated from sputum showed a resistance higher by 12-34% and 6-17% than those isolated from other specimens, such as pharyngeal exudate, where the resistance to rifadin was lower by 10%. beta-lactamases were present in 27.7% of the strains: H. parainfluenzae--36%, H. paraphrohaemolyticus--25%, H. influenzae--17.8% and H. parahaemolyticus--15.7%; in strains from sputum--34.2%, pharyngeal exudate--28.8% and from other specimens--6.6%. No correlations were noticed between the biotype and the clinical manifestation or the resistance to the antibiotic, a higher frequency of beta-lactamase production being reported in H. influenzae biotype V and H. parainfluenzae biotypes II and IV.     

Agglutination,Toxigenicity and Sorbitol Fermentation of Clostridium difficile

A total of 79 Clostridium difficile strains from different sources (50 strains from the fecal specimens of healthy adults, 13 from patients receiving antibiotics without gastrointestinal complications, 13 from antibiotic-associated pseudomembranous colitis (PMC) or diarrhea patients, and three strains from ATCC) were investigated for agglutinability, using formol-treated cells as antigen, in relation to toxigenicity. C. difficile strains tested were divided into four serovars, I, II, III, and IV, by the cross-agglutination test. The agglutinin absorption test revealed that strains of serovar I, agglutinable with high titers (5,120–10,240) to antiserum prepared against a highly toxigenic C. difficile strain, ATCC 17859, possessed the serovar-specific antigen. All of the strains of serovar I were highly toxigenic and all 13 strains isolated from the fecal specimens of antibiotic-associated PMC or diarrhea patients belonged to this serovar, whereas 19 (38%) out of 50 strains from healthy adults and four (30.8%) out of 13 strains from patients receiving antibiotics without gastrointestinal complications possessed this antigen. None of the strains of other clostridial species than C. difficile were agglutinated by the three reference antisera used. Further study on the sugar fermentation test disclosed that the sorbitol-fermenting property of C. difficile is very closely related to the toxigenicity and agglutinability.     

Tuberculin Reactions in Bulls And Boars Sensitized with Atypical Mycobacteria from Sawdust

Due to a planned export from a combined bull and boar station, more than 70 boars and 100 bulls were examined by tuberculin tests. Distinct reactions to avian tuberculin appeared in about half of the animals. Many of them also reacted to bovine tuberculin. For diagnostic purposes, many of the reactors were slaughtered, and samples from these and from the environment were examined bacteriologically. Strains of Mycobacterium avium were isolated from only 2 out of 14 reacting boars and from none of the 23 reacting bulls. No isolation of Mycobacterium bovis was made. However, atypical mycobacterial strains, classified as Runyon Group III and IV, were isolated from 3 boars, 2 bulls, 1 pigeon and from many samples of sawdust. The isolation of identical fast-growing mycobacterial strains from the sawdust used in the pens for the reacting boars and bulls, was especially remarkable. The strains differed enzymatically and biochemically from those isolated from other sources. This indicated a possible sensitization of the animals with similar mycobacterial strains. Possible cross-reactions to avian and bovine tuberculin were investigated in tuberculin assays with guniea pigs and pigs sensitized to one of the mycobacterial strains isolated. Distinct reactions to both avian and bovine tuberculin appeared in all the animals. From these results it was concluded that the tuberculin reactions in the boars and the bulls were not due to any tuberculous infection in the herd, but to a sensitization of the animals with atypical mycobacteria.     

Genetic and environmental factors affecting mating type frequency in natural isolates of Tetrahymena thermophila

In Tetrahymena thermophila mating type alleles specify temperature sensitive frequency distributions of multiple mating types. A-like alleles specify mating types I, II, III, V and VI, whereas B-like alleles specify mating types II through VII. We have characterized the mating type distributions specified by several A- and B-like genotypes segregated by genomic exclusion from cells isolated from a pond in northwestern Pennsylvania. The B-like genotypes are alike in specifying very low frequencies of mating type III, but differ with respect to the frequencies of other mating types, particularly II and VII. An A-like genotype specifies a high frequency of mating type III and is unstable in successive generations for the expression of mating type II, suggesting a possible modifier. Inter se crosses performed at 18 degrees C, 28 degrees C and 34 degrees C showed that each genotype specifies a frequency distribution that is uniquely affected by temperature. No mating type was affected the same way by temperature in all genotypes. In A/B heterozygotes, the B-like genotype exhibited partial dominance. The genotypes described here differ significantly from previously described genotypes from the same pond, indicating that there are numerous mating type alleles. For frequency-dependent selection to equalize mating type frequencies, it must act not only on complex multiple alleles but also on the response of mating type alleles to temperature.     

Integron-bearing methicillin-resistant coagulase-negative staphylococci in South China, 2001–2004

A total of 53 methicillin-resistant coagulase-negative staphylococci strains isolated in a hospital in Guangzhou, China, were analyzed to detect class 1 integrons and SCC mec typing. Thirty strains had the class 1 integrase ( intI1 ) gene and 26 strains possessed the 3' conserved region of qacE Δ 1 - sul1 . Four different types of gene cassette arrays were found and a highly prevalent array of dfrA12-orfF-aadA2 gene cassettes was observed. Thirty class 1 integron-positive coagulase-negative staphylococci strains were subjected to Southern hybridization analysis; the result showed that class 1 integrons were located on chromosome, not plasmid. According to the results of SCC mec typing for 30 integron-bearing MRCNS strains, five, 15 and five strains belonged to type I, II and III SCC mec , respectively, and five strains were untypeable. For 23 non-integron-bearing methicillin-resistant coagulase-negative staphylococci strains, four, nine and seven strains belonged to type I, II and III SCC mec , respectively, and three strains were untypeable. None of the strains belonged to type IV or V. Twenty-three coagulase-negative staphylococci isolates of three Staphylococcal species that contained the dfrA12-orfF-aadA2 gene cassette array were phylogenetically unrelated to each other by randomly amplified polymorphic DNA, indicating that the gene cassettes might be disseminated in the clinical strains by a horizontal gene transfer.     

Integron-bearing methicillin-resistant coagulase-negative staphylococci in South China, 2001-2004.

A total of 53 methicillin-resistant coagulase-negative staphylococci strains isolated in a hospital in Guangzhou, China, were analyzed to detect class 1 integrons and SCCmec typing. Thirty strains had the class 1 integrase (intI1) gene and 26 strains possessed the 3' conserved region of qacEDelta1-sul1. Four different types of gene cassette arrays were found and a highly prevalent array of dfrA12-orfF-aadA2 gene cassettes was observed. Thirty class 1 integron-positive coagulase-negative staphylococci strains were subjected to Southern hybridization analysis; the result showed that class 1 integrons were located on chromosome, not plasmid. According to the results of SCCmec typing for 30 integron-bearing MRCNS strains, five, 15 and five strains belonged to type I, II and III SCCmec, respectively, and five strains were untypeable. For 23 non-integron-bearing methicillin-resistant coagulase-negative staphylococci strains, four, nine and seven strains belonged to type I, II and III SCCmec, respectively, and three strains were untypeable. None of the strains belonged to type IV or V. Twenty-three coagulase-negative staphylococci isolates of three Staphylococcal species that contained the dfrA12-orfF-aadA2 gene cassette array were phylogenetically unrelated to each other by randomly amplified polymorphic DNA, indicating that the gene cassettes might be disseminated in the clinical strains by a horizontal gene transfer.     

Associations of scrotal circumference with semen traits in young beef bulls

Associations of percent live sperm, sperm number, sperm concentration, sperm motility, and potential breeding efficiency score with scrotal circumference were studied using data from the records of 465 Polled Hereford and 264 Simmental bulls. Data were subdivided by breed and within breed into three groups according to scrotal circumference (SC) size. Group I bulls had SC measurements <32 cm. Group II included bulls with SC measurements between 32 cm and the mean measurement of the breed (i.e., 35 cm for Polled Hereford and 38 cm for Simmentals). Group III was comprised of bulls with SC values above their respective breed averages. Age and body weight differences were also evaluated in the three groups. Average age differences among bulls in the three groups differed by not more than nine days in the two breeds. Average body weight differences among the three groups were statistically significant (P<0.05) with the maximum difference between any two groups within breed being 80 pounds. Average differences in percent live sperm and sperm number were greater (P<0.05) in Groups II and III than in Group I. For sperm concentration ratings, sperm motility ratings and potential breeding efficiency score, and for both breeds, Group I had a significantly lower (P<0.05) number of bulls classified satisfactory and a significantly greater (P<0.05) number of bulls classified unsatisfactory than did groups II and III.     

The Relationship of Capsular-type of Staphylococcus epidermidis to Virulence and Induction of Resistance in the Mouse

Biochemical, immunological and morphological properties of mouse virulent Staphylococcus epidermidis strains ATCC-31432, SE-360 and SE-10 isolated from clinical specimens were compared. Heat-killed organisms and cell surface polysaccharides extracted from cell surface fractions induced resistance in mice only against challenge with the homologous strain. Hyperimmune rabbit serum prepared with these strains passively protected mice against challenge infection only with the homologous strain. Protective activity in immune sera was absorbed by homologous whole cell and cell surface polysaccharide, but not by heterologous organisms and cell surface polysaccharide. In agar diffusion tests, cell surface polysaccharides from strains ATCC-31432, SE-360 and SE-10 produced single precipitin lines only with the homologous antiserum. The outermost layer of ultra-thin sections of the three strains was labelled by homologous but not by heterologous ferritin-conjugated serum. Biochemical analysis of the cell surface polysaccharides showed that they were composed of hexosamine, glycerol, phosphorous, alanine, glycine and phenylalanine. The three strains ATCC-31432, SE-360 and SE-10 were regarded as different from each other. Thirteen of 300 fresh isolates of Staph. epidermidis randomly selected from human clinical specimens proved to be virulent for mice. With ATCC-31432, SE-360 and SE-10 tentatively designated as capsular-type I, type II and III, respectively, a majority of mouse virulent strains belonged to capsular-type II.     

Phenotypic differentiation of bifidobacteria of human and animal origins.

The phenotypes of 153 strains belonging or related to the genus Bifidobacterium were studied. These organisms included 38 collection strains and 115 wild strains (41 strains of human origin, 56 strains of animal origin, and 18 strains obtained from rivers or sewage). Our phenotypic analysis revealed seven main groups that were subdivided into 20 subgroups. Seven subgroups contained no type or collection strain. Among the human strains, the type strains of Bifidobacterium pseudocatenulatum and B. catenulatum fell into group I, which contained the type strains of B. adolescentis (subgroup Ib), B. dentium (subgroup Ic), and B. angulatum (ungrouped). The type strain of B. breve belonged to subgroup IIIa1, and the type strains of B. infantis and B. longum fell into subgroup IIIb1. Group VII comprised only wild strains that were isolated from human infant feces. Among the animal strains, group II consisted mainly of bifidobacteria that were isolated from pig feces and contained the type strains of B. suis (subgroup IIb), B. thermophilum (subgroup IIf), B. choerinum, and B. boum (ungrouped). Wild strains belonging to group V were isolated from pig, calf, cow, and chicken feces; this included the type strains of B. animalis (subgroup Va), B. magnum (subgroup Vb), B. pseudolongum, and B. globosum (subgroup Vc). The strains of human origin (groups I, III, and VII) were well separated from the animal strains (groups II, IV, and V). It was not surprising that the wild strains isolated from surface water or sewage were distributed in the animal groups as well as the human groups. Thus, bifidobacteria can be considered to be successful indicators of human or animal fecal pollution when they are correctly classified. The acidification patterns were not adequate to differentiate Bifidobacterium species, as determined previously by Mitsuoka (Bifidobacteria Microflora 3:11-28, 1984) and Scardovi (p. 1418-1434, in P. H. A. Sneath, N. S. Mair, M. E. Sharpe, and J. G. Holt, ed., Bergey's Manual of Systematic Bacteriology, vol. 2, 1986). However, enzymatic tests furnished new taxonomic criteria for the genus.     

OESTROUS BEHAVIOUR, OVULATION AND THE FORMATION OF THE CORPUS LUTEUM IN THE AFRICAN ELEPHANT,

A female elephant was observed throughout most of oestrus, and was then shot. The cow was mated by many different bulls during the course of oestrus. Initially, there seemed to be little competition between bulls for the cow, but as oestrus wore on, fighting broke out amongst the bulls and one eventually appeared to establish mastery, driving the other bulls away. It was interesting that this master bull showed no discharge from his temporal glands.     

Effect of mating stimuli on LH release in bulls

Fourteen crossbred beef bulls were assigned at random to receive one of four sexual stimuli treatments. Treatments consisted of: (1) controls (four bulls), no visual or physical contact with any cows; (2) false mount (two bulls), allowed to mount an estrual cow; (3) mated (four bulls), allowed to mount an estrual cow with intromission and ejaculation; (4) electroejaculated (four bulls), no exposure to cows. Serum from blood samples taken at 15-min intervals from -15 min to 2 hr from sexual stimuli were radioimmunoassayed for luteinizing hormone (LH). Four bulls had elevated levels at the pretreatment bleeding, but none of the stimuli induced or were associated with LH releases. Basal levels of LH were consistent within bulls but varied considerably among bulls. Conclusion is that stimuli associated with mating do not cause a release of LH.     

Some Properties of the Pili of Corynebacterium renale

Some properties of the pili of the gram-positive bacteria Corynebacterium renale were described. A relationship was found between the morphological features of pili and the types of C. renale. Strains of types I and III usually possessed a small number of pili, whereas those of type II possessed numerous pili. Thick and long bundles of pili characteristic of C. renale were frequently observable in type II strains. Piliation of C. renale was stable under various cultural conditions. No ability to agglutinate red blood cells was demonstrated by piliated strains of C. renale. Pili were isolated from the cells of C. renale and studied serologically by immunodiffusion. The pili of a type II strain were serologically identical with the pili of another type II strain but not with those of the strains belonging to types I and III. The pili were serologically distinct from the cell wall. The pili were broken into short pieces by boiling, but their antigenicity was increased after boiling.     

Behaviour and mating performance of paddock-mated beef bulls

Paddock-mating performance was studied in seven beef bulls which were each joined to 28 or 29 females of mixed dairy and beef breeds for 8 weeks in single-sire joining groups in paddocks ranging from 4.0 to 28 ha. Seven unmated bulls were used as control animals. The estimated weight of testicular tissue per cow joined ranged from 12 to 32 g. Liveweight and testicular size and tone were recorded prior to, during and after joining. There was no apparent effect of mating on testicular characteristics of bulls, and pre-joining testicular size of the bulls was not significantly related to the calving results of the cows.Mating activity of the bulls was detected with a chin-ball harness and by behavioural observations covering eight intervals during the joining period. During the first 21 days (cycle I) of the joining period, 81% of the cows were mated and this increased to 89% by the end of the joining period. The overall return rate was 21%, the overall calving rate was 85% and no significant differences between the seven bulls were recorded in these two parameters. The relatively high level of sexual activity during cycle I apparently had no adverse effect on the fertility of the bulls since there was an even distribution over time of the return to first service of cows mated during cycle I. Mating activity of the bulls during cycle I had no significant effect on the mean percentage of time spent grazing or lying, but time spent standing was lower (P < 0.001) during cycle I than the remainder of the joining period.General activity, expressed as km travelled per day, was measured at regular intervals during the joining period for one working bull and one heifer in each of four mating groups. Mean activity of the bulls was significantly higher (11.3 km/day) during cycle I than the remainder of joining (2.45 km/day) and was significantly correlated (r = 0.8) to the mean number of cows mated per bull per day.     

Protein G genes: structure and distribution of IgG-binding and albumin-binding domains

Protein G (also designated Fc receptor type III) is the IgG-binding protein of group C and G streptococci. Protein G has also been shown to bind human serum albumin but at a site that is structurally separated from the IgG-binding region. From the known gene sequence of protein G, two synthetic oligonucleotides were constructed for use as probes in DNA-hybridization experiments to study the structure and distribution of the albumin- and IgG-binding regions in bacterial strains belonging to different species. Thus, one of the probes corresponded to repeats within the IgG-binding region (I) and the other corresponded to repeats in the albumin-binding encoding region (II). Probe I showed strong hybridization to DNA isolated from 31 human group C and G strains, whereas hybridization to probe II was variable. With the three restriction endonucleases used, three restriction patterns were found in Southern blot experiments. No fundamental difference could be detected in hybridization experiments, either between strains of group C and G streptococci, or between isolates of different clinical origin. No hybridization to DNA from other bacterial species was found.     

Phosphorylation of the EGF receptor from A431 epidermoid carcinoma cells by three distinct types of protein kinase C

Three distinct types of protein kinase C obtained from rat brain cytosol phosphorylated the EGF receptor of A431 epidermoid carcinoma cells at different rates. This receptor was phosphorylated most rapidly by type III protein kinase C, but slowly by type I enzyme. Type II enzyme showed intermediate activity. Chromatographic analysis indicated that A431 cells possessed only one of the three types found in rat brain, which apparently corresponded to type III enzyme. This type of protein kinase C, that is encoded by the alpha-sequence or a closely related sequence, appeared to be expressed commonly in many tissues and organs. The result implies that type III enzyme may play roles in growth promotion.     

Proteins of the accessory sex glands associated with the oocyte-penetrating capacity of cauda epididymal sperm from holstein bulls of documented fertility

We previously reported that accessory sex gland fluid (AGF) from high fertility (HF) bulls influenced the oocyte-penetrating capacity of cauda epididymal sperm from low fertility (LF) bulls, based on in vitro fertilization (IVF) assays. The present study determined if AGF proteins were associated with these effects. Nineteen IVF assays with 12 bulls were grouped as follows. Group I (n = 8): assays where sperm from LF bulls exposed to AGF from HF bulls had greater oocyte penetration than exposed to homologous AGF. Group II (n = 7): sperm from LF bulls to AGF from HF bulls versus homologous AGF showed no significant differences. Group III (n = 4): sperm from LF bulls treated with homologous AGF had greater fertility than sperm treated with AGF from HF bulls. Sire fertility was based on nonreturn rates (NNR) and AGF collected by artificial vagina from bulls with cannulated vasa deferentia. Two-dimensional SDS-PAGE maps of AGF were analyzed by PDQuest and proteins identified by tandem mass spectrometry and Western blots. Differences in spot intensity between AGF of HF and LF bulls were compared across groups of IVF assays (P < 0.05). The expression of BSP A1/A2 and A3, BSP 30 kDa, clusterin, albumin, phospholipase A(2) (PLA(2)), and osteopontin was greater in the AGF of HF bulls in Group I as compared to Groups II and III. Conversely, there was less nucleobindin in the AGF of HF bulls in Group I than in Groups II and III. This is the first report of nucleobindin (58 kDa/pI 5.6) in male reproductive fluids, using both immunoblots and mass spectrometry. Thus, the effect of AGF from HF bulls on epididymal sperm is likely the result of specific proteins expressed in the AGF.     

Infectious Bovine Rhinotracheitis Virus Infection in Bulls, with Special Reference to Preputial Infection

In an experiment with infectious bovine rhinotracheitis (IBR) virus in two bulls, observed over a period of 122 weeks, the pattern of virus release was studied. Recurrent, unprovoked release of virus was demonstrated after one year in a nasal washing from one of the bulls and in preputial washings of both on 13 and 4 occasions, respectively, and finally in weeks 113 and 110, although clinical disease was not observed. During periods of recurrent virus release, concentrations of virus in the prepuce were generally much lower than during the period of primary infection; usually, however, they were not of negligible titer. The frequency of such periods and the virus titers observed strongly suggest that an IBR antibody carrier should always be considered as a potential source of infection to other animals. When virus was demonstrated in semen an almost equal amount was found in the preputial washing (50 ml). In week 120, virus replication in the respiratory tract and prepuce was induced in both bulls by prednisolone injections. It is concluded that antibody carriers will rarely attain a state of absolute immunity.