Pathogenesis of porcine reproductive and respiratory syndrome virus infection in mid-gestation sows and fetuses.

Two experiments were undertaken to evaluate whether porcine reproductive and respiratory syndrome (PRRS) virus was able to cross the placenta and infect midgestation fetuses following intranasal inoculation of sows and whether PRRS virus directly infected fetuses following in utero inoculation. In experiment 1, eight sows between 45 and 50 days of gestation were intranasally inoculated with PRRS virus (ATCC VR-2332), and four control sows were inoculated with uninfected cell culture lysate. Virus inoculated sows were viremic on postinoculation (PI) days 1, 3, 5, 7 and 9, shed virus in their feces and nasal secretions, and became leukopenic. Sixty-nine of 71 fetuses from principal sows euthanized on PI day 7, 14 or 21 were alive at necropsy and no virus was isolated from any of the fetuses. Two principal sows that farrowed 65 and 67 days PI delivered 25 live piglets and three stillborn fetuses. The PRRS virus was isolated from two live piglets in one litter. In experiment 2, laparotomies were performed on five sows between 40 and 45 days of gestation and fetuses were inoculated in utero with either PRRS virus alone, PRRS virus plus a swine serum containing PRRS antibodies, or uninfected cell culture lysate. Three sows were euthanized on PI day 4 and two sows on PI day 11. Viral replication occurred in fetuses inoculated with virus alone and was enhanced in fetuses inoculated with virus plus antibody. No virus was isolated from control fetuses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation of a cytopathic virus from weak pigs on farms with a history of swine infertility and respiratory syndrome.

Severe clinical signs of swine infertility and respiratory syndrome (SIRS) of unknown cause were observed in several Minnesota swine farms between November 1990 and March 1991. Forty-five lung samples of weak pigs were collected from 13 swine farms, and virus isolation was attempted using swine alveolar macrophage (SAM) cultures. A cytopathic virus was isolated from 19 lung samples collected from 6 different farms. Four pregnant sows were infected intranasally with a tissue suspension from which virus was isolated, and 4 6-week-old pigs and 2 contact pigs were infected intranasally with 1 of the isolates. The 4 sows farrowed 12 stillborn and 32 normal pigs. Virus was recovered from 10 of 19 pigs examined. Infected 6-week-old pigs were clinically normal except for slightly elevated rectal temperatures and mild respiratory signs. No or mild interstitial pneumonic lesions were observed in inoculated pigs, but the lesion was obvious in the 2 contact pigs. Seroconversion was observed in sows and pigs as measured by indirect fluorescent antibody (IFA). Serologic identification of the isolates was carried out by IFA using reference serum prepared from an experimentally infected sow. A cytoplasmic fluorescence was observed on the SAM monolayers infected with each of the 19 different isolates. Fluorescence was also observed when the monolayers were tested with SIRS virus ATCC VR-2332-infected sow sera. Replication of the isolates was not affected in the medium containing 5-iodo-2'-deoxyuridine but was inhibited by treatment with ether. The isolates were relatively stable at 56 C and did not agglutinate with various erythrocytes tested.(ABSTRACT TRUNCATED AT 250 WORDS)     

Restriction endonuclease analysis of various strains of Mycobacterium paratuberculosis isolated from cattle

Deoxyribonucleic acid (DNA) preparations from 3 reference strains of Mycobacterium paratuberculosis and from 23 isolates of M paratuberculosis obtained from cattle in New Zealand were characterized by restriction endonuclease analysis, using the enzymes BstE II, Pvu II, and Bcl I. Patterns of DNA fragments for strain 18 (one of the reference strains) differed markedly from patterns of other strains, indicating genetic differences between strain 18 and the other strains of M paratuberculosis evaluated. The other 2 reference strains (TMC 1613 and Weybridge strain 316) and all but 1 of the isolates from cattle had identical patterns with the 3 enzymes. These 2 reference strains differed from each other in their dependence on exogenous mycobactin, but this was not reflected in their restriction patterns. The single variant isolate from cattle had patterns identical to those of the other isolates, using Pvu II and Bcl I, and had only 1 fragment line difference with BstE II. Although close genetic homogeneity of cattle strains of M paratuberculosis prevented development of a typing system on the basis of restriction endonuclease analysis, the results provided a basis for genomic comparison with other closely related organisms.     

Experimental infections of horses with Legionella pneumophila

Attempts to infect horses with Legionella pneumophila were undertaken to determine pathogenicity and to evaluate the possibility that horses serve as a reservoir for the organism. A previous study showed that the prevalence of antibodies to L pneumophila in the equine population exceeded 30% of over 600 sera examined. Horses were infected experimentally with the Philadelphia 1 or Bloomington 2 strain of L pneumophila IV or by aerosolization. Signs of clinical illness were restricted to a transient febrile response. A transient decrease in circulating lymphocytes occurred 2 days after inoculation. At necropsy, only moderate generalized lymphadenopathy was noted. Histologically, the lungs contained evidence of a low-grade inflammatory response characterized by focal proliferation of alveolar lining cells, with few neutrophils and eosinophils. Lymph nodes had evidence of reactive hyperplasia. The tissue response to Bloomington 2 strain was slightly more pronounced than that to Philadelphia 1. Attempts to reisolate L pneumophila from blood and nasal or pharyngeal swabs were unsuccessful. The organism was not isolated by culturing tissues obtained at necropsy, nor was it demonstrated by tissue-staining techniques. However, all horses exhibited a marked increase in agglutinating antibodies to L pneumophila serogroups (SG) 1 and 3 as early as 4 days after inoculation. The serologic response was confirmed by indirect immunofluorescence and was shown to consist predominantly of immunoglobulin M by 2-mercaptoethanol treatment. Agglutinating antibodies persisted at least 4 months after infection. On the basis of these studies, the pathogenicity of L pneumophila SG 1 and 3 for the horse appears to be low. There is no evidence to support a role for the horse in the maintenance of these organisms in nature. Horses may be exposed in the environment and maintain a relatively long-lived serologic response to L pneumophila. However, it is also possible that they become infected with other strains of L pneumophila or Legionella-like organisms more pathogenic for horses, or other non-Legionella bacteria, which elicit a cross-reacting serologic response to L pneumophila SG 1 to 4.     

The prevalence of Dirofilaria immitis in dogs in Sydney

Four hundred and four dogs from 9 pounds in Sydney were examined for circulating microfilariae and antigens of Dirofilaria immitis. One hundred of these were also examined post mortem for adult heartworms. The prevalence of infection in the 404 dogs as shown by serology was 11.4%, and 5.9% had circulating microfilariae of D immitis. Adult heartworms were present in 15 of 100 dogs. Dipetalonema reconditum microfilariae were present in 3.7% of dogs. Dirofilariosis is still a common and important parasite of dogs in the Sydney region and chemoprophylaxis is recommended.     

Genetic diversity and population structure of Brachiaria brizantha (A.Rich.) Stapf accessions from Ethiopia

Brachiaria is a tropical, warm-season grass native to Africa. It is an extensively cultivated forage in the tropics with proven benefits on livestock productivity. Brachiaria is well-known for high biomass production, animal nutrition, carbon sequestration, biological nitrification inhibition, soil conservation, and adaptation to drought and low fertility soils. However, the use of Brachiaria grass for fodder production in Africa has been little explored largely due to lack of cultivars suitable to different production environments. The exploration and use of natural diversity is fundamental for an efficient Brachiaria breeding program. We analysed genetic diversity and population structure of 112 Ethiopian Brachiaria brizantha accessions using 23 microsatellite markers. A total of 459 alleles were detected with an average polymorphic information content of 0.75 suggesting high discriminating ability of these markers. The molecular variance analysis showed a high contribution (86%) of within-cluster differences to the total variation. Three allelic pools revealed by STRUCTURE analysis in 112 accessions were in agreement with the clustering patterns seen in neighbor-joining tree and principal coordinates analyses. A core collection of 39 B. brizantha accessions was constituted. This study concludes a high genetic diversity of Ethiopian B. brizantha accessions and their importance in Brachiaria breeding programs.     

Epidemiological investigation of Leptospira spp. in a dairy farming enterprise after the occurrence of three human leptospirosis cases

An epidemiological investigation was conducted in an unvaccinated dairy farming enterprise in which three workers on one of the milking herds (Herd 1) were diagnosed with leptospirosis due to serovars Hardjo (H) (n = 2) and Pomona (P) (n = 1) between January and March 2015. Blood and urine samples were collected from milking cows in Herd 1 (N = 230) and Herd 2 (N = 400), rising one‐ (R1, N = 125) and rising two‐year‐old (R2, N = 130) replacement heifers, and four pigs associated with Herd 1, in March 2015. Sera were tested using the MAT for serovars H, P, Copenhageni (C), Ballum (B) and Tarassovi (T), and urine samples were tested by qPCR. Seventy‐five per cent of 109 cows in Herd 1 and 36% of 121 in Herd 2 were seropositive (≥48), predominantly to H and P, and 23% of 74 cows in Herd 1 and 1% of 90 cows in Herd 2 were qPCR positive. Fifty‐five per cent of 42 R2 heifers were seropositive to T. No R1 and 17% of 42 R2 heifers were qPCR positive. Subsequently, all cattle were vaccinated for H and P, and Herds 1 and 2 were given amoxicillin. After the booster vaccination, 7% of 91 in Herd 1, 2% of 82 in Herd 2 and 11% of 38 R1 heifers (sampled as R2) were PCR positive. After the amoxicillin treatment, no cows in Herd 1 and 5% of 62 cows in Herd 2 were urine PCR positive. Calves and pigs were seropositive to H, P, C and B. Vaccination and antibiotic treatment appeared effective in reducing the risk of exposure of workers to vaccine serovars. However, evidence of non‐vaccine serovars indicated that workers likely remain at risk of exposure to Leptospira.     

Vaginal isolation of beta‐haemolytic Streptococcus from bitches with and without neonatal deaths in the litters

The aim of the study was to identify beta‐haemolytic streptococci in the vagina of bitches who had delivered healthy litters and bitches who had delivered litters in which neonatal deaths occurred. Fifty‐one bitches divided into two groups were used. Group 1 (G1) included 28 bitches that had delivered healthy litters and group 2 (G2) included 23 bitches that had delivered puppies who died in the neonatal period. Two vaginal samples were taken, one in proestrus and the other at the end of gestation (EG). Beta‐haemolytic Streptococcus (BS) was isolated from 16 bitches (57%) in G1 and from 21 bitches (91%) in G2. The bacteriological cultures, serological tests (Streptex^®) and PCR assay allowed identification of Streptococcus canis and Streptococcus dysgalactiae in G1 and G2. Ultramicroscopic studies allowed the observation of M Protein and capsules in strains of S. dysgalactiae and S. canis in G1 and G2. The S. canis strains isolated from G2 showed thicker capsules than S. canis strains isolated from G1 (234 ± 24.2 vs 151.23 ± 28.93 nm; p < .001.). No differences were observed in capsule thickness between strains of S. dysgalactiae isolated from G1 and G2 (210 ± 13.54 vs 211.66 ± 19.67 nm; p > .70). All strains of beta‐haemolytic Streptococcus isolated were penicillin sensitive. Penicillin was administered from EG to 5 days post‐partum in 10 G2 females with isolation of BS (G2A). Saline solution was administered in eleven G2 females with isolation of BS (G2B). Ninety per cent of the puppies survived in G2A and 25% survived in G2B. Our results suggest BS is involved in canine neonatal deaths.