Cultural and biochemical characterization of Actinobacillus and Actinobacillus-like species from ram lambs with epididymitis

Cellular, colonial, cultural, and biochemical characteristics of 25 field strains of gram-negative pleomorphic bacilli from rams with epididymitis were compared with Actinobacillus actinomycetemcomitans American Type Culture Collection (ATCC) strain 29522 and Actinobacillus seminis ATCC strain 15768. Three field strains were identified as A. actinomycetemcomitans, 15 as A. seminis, and 2 as Haemophilus agni; however, 5 strains (3 in group A and 2 in group B) were not identified as species in the genera Actinobacillus, Haemophilus, or Pasteurella based on the taxonomic criteria in Bergey's manual of systematic bacteriology. The 5 Actinobacillus-like organisms in groups A and B were predominantly gram-negative coccobacilli and exhibited less pleomorphism than the 2 Actinobacillus species. The colonial morphologies of groups A and B were similar to the 2 Actinobacillus species but were smaller in diameter and had a pale yellow color. Groups A and B, like the actinobacilli, were facultative anaerobic and capnophilic, did not grow on MacConkey agar, and were catalase-positive and oxidase-positive. Group A reduced nitrate but group B did not. The A. seminis strains utilized ornithine, and group A utilized arginine; but group B did not utilize either ornithine or arginine. All strains failed to utilize lysine or tryptophane. All strains produced acid but no gas from glucose, and the utilization of other carbohydrates varied markedly both between and within the 5 groups of bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)     

Unilateral orchitis and epididymitis caused by Salmonella enterica subspecies diarizonae infection in a ram.

A case of unilateral suppurative epididymo-orchitis associated with Salmonella enterica subsp. diarizonae serovar 61:k:1,5,(7) infection is described in a 2-year-old ram. Gross lesions were characterized by severe enlargement of the scrotal contents, fibrous adhesions between testicular layers, coexistence of epididymal abscesses and foci of fibrinous exudate, and testicular atrophy. Microscopically, testicular and epididymal microabscesses and diffuse inflammatory infiltrates with abundant macrophages containing short Gram-negative rods were observed. Superimposed on the chronic lesions were fibrin deposits with clusters of neutrophils, as well as walled-off granulation tissue. Bacterial colonies were also identified in thrombosed spermatic cord vessels, scrotal lymph nodes, lung, and liver. S. enterica subsp. diarizonae serovar 61:k:1,5,(7) was isolated from the affected testis. To the authors' knowledge, this is the first report of infection of the testis and epididymis by Salmonella in rams. This organism must be taken into account in the differential diagnosis of ovine genital infections.     

Production and preliminary characterization of monoclonal antibodies to Actinobacillus sp isolated from epididymitis lesions in a ram

A panel of 55 monoclonal antibodies to an Actinobacillus sp isolate (As8C strain) cultured from the epididymides of an infected ram was produced. Cell lines producing 5 of these antibodies were cloned and expanded by hybridoma tumor production in Balb/c mice. An isotype profile revealed that 1 cloned antibody belonged to the immunoglobulin (Ig) M class and the 4 remaining antibodies belonged to the IgG class. Within the IgG class, 1 clone produced IgG1, 1 clone produced IgG2a, and 2 clones produced IgG2b. Ascites fluid antibody titers from the cloned hybridomas ranged from 6,400 to 51,200, as determined by enzyme-linked immunosorbent assay. Specificity of the antibodies to target As8C antigens could be demonstrated by enzyme-linked immunosorbent assay inhibition. Ascites fluid from 2 clones contained antibodies that agglutinated As8C. Two additional clones produced antibodies capable of only partial agglutination, whereas 1 clone produced antibody that did not agglutinate As8C. The indirect fluorescent antibody test revealed that target antigens for at least 4 of the 5 monoclonal antibodies were most likely located on the bacterial cell surface. Antigens were extracted from As8C, using 5 surface active chemicals. An attempt to immunoprecipitate these antigens in agarose by reacting individual extracts with each of the antibodies was unsuccessful.     

Comparison and partial characterization of the protein profiles and outer membrane antigens of Actinobacillus species isolated from ram lambs with epididymitis

Three monoclonal antibodies (LG17, LG30, LG33) were used in the indirect fluorescent antibody test, the ELISA, and the immunoelectrotransfer blot technique to identify group-specific and strain-specific epitopes on the outer membranes of Actinobacillus seminis, A actinomycetemcomitans, and 17 field isolates of Actinobacillus spp. The field isolates had been obtained by bacteriologic culture of specimens from ram lambs with epididymitis. Only antibody LG33 consistently had specificity for an outer membrane epitope shared by most of the bacterial isolates tested. Staining of polyacrylamide gels with periodic acid-Schiff reagent, Sudan black B, and Coomassie brilliant blue R250 indicated that target antigens for antibodies LG17 and LG33 contained carbohydrate and lipoprotein components, respectively. The chemical composition of the LG30 target antigen was not determined because of its instability after exposure to sodium dodecyl sulfate. Discontinuous-gradient polyacrylamide gel electrophoresis in sodium dodecyl sulfate and spectrophotometric scans of the gels were used to analyze n-octyl-beta-D-glucopyranoside protein extracts from A seminis, A actinomycetemcomitans, and 13 representative field isolates of Actinobacillus spp. Bacterial isolates could be grouped according to their protein profiles. The first group consisted of A seminis, A actinomycetemcomitans, and 7 field isolates of Actinobacillus spp, all of which shared common protein bands with molecular masses of approximately 94 kilodaltons (kD), 64 kD, 60 kD, 52 kD, 44 kD, and 26 kD. The second group was composed of 6 field isolates, each with unique protein profiles; isolates had relatively few protein bands in common. These data suggested that members of the genus Actinobacillus cultured from ram lambs with epididymitis probably include a number of various strains.     

Prevalence, causes and laboratory diagnosis of subclinical mastitis in the goat

Five goat herds were examined to determine the prevalence and causes of subclinical mastitis and to assess the value of some laboratory tests currently used on milk samples as aids in the diagnosis of caprine mastitis. In the 170 samples taken from the pairs of mammary glands of 85 goats, the prevalence of infection in the different herds ranged from 15 per cent to 79 per cent of halves. Just over one-third (36 per cent) of all halves were infected, the organisms isolated being coagulase-negative staphylococci (80 per cent), coagulase-positive staphylococci (16 per cent), alpha-haemolytic streptococci (2 per cent) and Pasteurella haemolytica (2 per cent). Neither anaerobic organisms nor mycoplasmas were found. Tests confirmed that the coagulase-positive staphylococci were pathogens but that the coagulase-negative staphylococci rarely caused detectable disease in the caprine udder. The large between-herd variation in the geometric mean cell counts of uninfected milk samples means that somatic cell counts, the Whiteside test and the California mastitis test, are unreliable as aids in the diagnosis of caprine mastitis.     

Bacterial epididymitis in two stallions

Two stallions had unilateral bacterial epididymitis attributable to S zooepidemicus infection. Diagnosis was based on bacterial isolation, WBC in the semen, higher than normal blood fibrinogen concentration, and leukocytosis with regenerative left shift. One horse had high seminal pH. Ultrasonography of the involved epididymides revealed changes consistent with the appearance of abnormal accumulation of exudate in the tail of the epididymis. Treatment included unilateral orchiectomy and antibiotic administration. In stallion 1, the infection persisted despite treatment. Treatment result was not available for stallion 2.     

Effects of ram preexposure and ram breed on fertility of ewes in summer breeding

Suffolk x Rambouillet ewes were used in two consecutive years to test the ability of Dorset and Suffolk yearling rams to stimulate ovulation and estrus in early summer (late June and July) breeding. After spring weaning about June 1, ewes were randomly divided into three groups and for 2 wk either were isolated from rams or were preexposed to either yearling Dorset or Suffolk rams by penning of rams in proximity to ewes. Ewes then were rerandomized into two groups for joining with either yearling Dorset or Suffolk rams fitted with marking harnesses. Serum progesterone profiles and crayon marks were used as indicators of ovulation and estrus. Overall lambing rate was not different among preexposure treatments for the 2 yr. However, early lambing rate (in the first 14 d of lambing season) was significantly affected by preexposure group (24% for Dorset vs 9% for Suffolk and 10% for isolation), especially in the 2nd yr (31% for Dorset vs 8% for both Suffolk and isolation). Ewes preexposed to Dorsets also lambed 10 d earlier (P less than .05) than ewes preexposed to Suffolks in yr 2, but the groups did not differ in yr 1. Ewes exposed to Dorset rams for breeding had higher overall lambing rates than ewes bred to Suffolks (75% vs 54%), especially in the 1st yr (77% vs 48%). Lambs sired by Dorset rams were born 6 d earlier in yr 1 (P less than .05) but only 1 d earlier in yr 2. Data from both years indicate that Dorsets are superior to Suffolks in ability to sire lambs in fall lambing systems.     

Serodiagnosis of Histophilus ovis-associated epididymitis in rams

An ELISA for the detection of antibodies to Histophilus ovis was used to evaluate the association of epididymal lesions in rams with serologic response to His ovis. Comparison of ELISA results for His ovis in groups of rams with epididymal lesions with ELISA results of clinically normal rams (control group) revealed a significant difference (P less than 0.01) between the control group and those rams from which His ovis was isolated. A significant difference (P less than 0.01) was noticed between the control group and rams with lesions from which an organism other than His ovis or Brucella ovis was isolated. Additionally, a significant difference (P less than 0.01) was noticed in ELISA results between the control group and affected rams from which no organism was recovered and in which the epididymal lesion was not limited to the head of the epididymis. A difference was not detected in the His ovis ELISA results between control rams and rams with lesions associated with a B ovis infection or rams from which no organism was recovered and in which the epididymal lesion was limited to the head of the epididymis. The serologic findings in our study suggest that His ovis is more important in the development of epididymitis in rams than culture results alone would indicate.     

Dystocia in Friesian mares: Prevalence,causes and outcome following caesarean section

Friesian horses appear to suffer a relatively high incidence of dystocia but there are, to the authors' knowledge, no published reports on the incidence or types of dystocia in Friesian horses. A retrospective survey of clinical details and post treatment recovery was performed for 66 mares referred to Wolvega Equine Hospital for dystocia during 2001–2006. Friesian mares appear to be particularly prone to dystocia due to fetal ankylosis or transverse presentation. However, despite dystocia of relatively long duration prior to caesarean section, post operative survival and fertility of mares and, more surprisingly, survival of foals were similar to those reported in surveys for other breeds.     

Prevalence and major bacterial causes of bovine mastitis in Asella,South Eastern Ethiopia

A cross sectional study was conducted in and around Asella town from November 2007 to April 2008 on dairy cows to determine the prevalence of mastitis, impact of risk factors and isolate the dominant mastitis causing bacteria on total of 223 lactating cows, of which 92 were indigenous Arsi, and 131 Holstein Zebu cross by using clinical examination and California mastitis test (CMT). Of these 144 (65.6 %) were positive by clinical examination and CMT for clinical and sub clinical mastitis, with prevalence of 26.5 % and 38 %, respectively. There was a significant difference (P < 0.05) on the prevalence of mastitis between cows kept under different hygiene of milking process. Similarly a significant difference on the prevalence of mastitis between the two breeds (P < 0.05) was also observed. From 144 CMT and clinically positive milk samples analyzed microbiologically, 133 were culturally positive for known mastitis pathogens and while 11 were negative. The dominant bacterial isolates in the study animals were Staphylococcus species (41.4 %), Streptococcus species (24.8 %), and other Gram positive rods and Gram negative enteric bacteria (33.8 %). Good hygiene in milking process, milking clinically infected cows at last, culling chronic mastitis carriers, treating clinically infected cows and dry period therapy could reduce the prevalence of contagious mastitis in the study area.     

Ultrasonographic detection of chronic epididymitis in a stallion

Clinical signs of chronic epididymitis in a stallion included recurrent signs of inguinal pain--retraction of the right testis into the inguinal area, resentment of palpation of the right testis, and right-sided scrotal swelling. The tail of the right epididymis was firm and seemed to be adhered to the testis. The tail of the left epididymis felt firmer than normal. Ultrasonographically, the diameter of the head of the epididymis was considered larger than normal, was hyperechoic in relation to the right testis, and had an irregular border. Several bright 1- to 5-mm-diameter echogenic areas that alternated with less echogenic areas were seen in the head of the right epididymis. Ultrasonographic findings were interpreted as fibrosis attributable to chronic inflammation. Unilateral castration (right-sided) was performed, but the prognosis was guarded because of presumed involvement of the left epididymis.