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PubMed2000

Effects of porcine reproductive and respiratory syndrome virus (isolate tw91) on porcine alveolar macrophages in vitro.

To verify the role of porcine reproductive and respiratory syndrome virus (PRRSV) infection on pulmonary defense mechanisms, alterations in the viability, morphology, and various functions of porcine alveolar macrophages (AMs) were evaluated in vitro for 2-72 h after exposure to a Taiwan isolate, tw91, at a multiplicity of infection (MOI) of 0.1. A low but constant rate of infection, around 5%, was seen in AMs from the PRRSV-infected group throughout the study. When compared with a mock-infected group, AMs from the PRRSV-infected group had a significantly lower viability at 18-72 h post-infection (HPI) as determined by trypan blue dye exclusion. Also during this time period, the cells showed morphological changes, including rounding, bleb formation, and rupture. The phagocytic and microbicidal capacity of AMs against Candida albicans was significantly inhibited after 6 HPI. Although the total amount of superoxide anion (O2-) and hydrogen peroxide (H2O2) produced by the AMs was reduced after 18 and 12 HPI, respectively, the amount of production was enhanced in both reactive oxygen species on a per viable cell basis after 12 HPI. In contrast, the level of bioactive tumor necrosis factor alpha (TNF-alpha) secretion, either total or on a per viable cell basis, was markedly reduced soon after PRRSV infection, up to 36 HPI, followed by a rebound thereafter. Prostaglandin E2 (PGE2) production was enhanced, both in total and on a per viable cell basis, in the first 6 h of infection, especially at 2 HPI. However, it became lower than that of the control after 36 HPI. The results indicated that PRRSV infection could cause, directly and/or indirectly, not only death of AMs but also adverse alterations in their morphology and function, although some of the effects seemed to be reversible. Because AMs are crucial to the host against airborne pathogens, PRRSV infection may potentially predispose pigs to secondary pulmonary infections.

Animals↗
PubMed2002

Leader-body junction sequence of the viral subgenomic mRNAs of porcine reproductive and respiratory syndrome virus isolated in Taiwan.

In combination of utilizing a leader specific primer and primers complementary to porcine reproductive and respiratory syndrome virus (PRRSV) genome through RT-PCR, the leader junction sequences of subgenomic mRNA (sg mRNA) was identified from a Taiwanese isolate of PRRSV. Thirty-six cDNA clones derived from sg mRNAs 2, 3, 4, 5, 6 and 7 were determined. The junction sequences analyzed from different sg mRNA were found to contain a similar 5 nucleotide sequence motif, (U/G)(C/A)(A/G)CC. The distance between the junction site and the translation initiation codon of the down stream open reading frame varied from 4 to 226 nucleotides. Minor heterogenecity was observed in the nucleotide sequence surrounding the junctions from all six sg mRNA analyzed. However, for sg mRNA 7, two junction sites approximately 103 nucleotides apart from each other were identified. The additional site is a new junction not previously reported in sg mRNA 7 from other PRRSV strains.

Base Sequence↗
PubMedDate not supplied

[Reproductive behavior and longevity of isolated and grouped couples of Lucilia cuprina, under controlled conditions].

The Lucilia cuprina laboratory breeding depends, fundamentally, on reproductive behavior and longevity information. In the present study it was verified that, due to the species isolation, there was a remarkable increase in the number of eggs/female (1344.18), when compared to the number of grouped eggs/female (732.78). The eggs viability was reduced due to the isolation of L. cuprina female. The longevity rate was estimated in 29.41 and 31.20 days for grouped males and females, respectively. For isolated males and females the longevity rate was 25.20 and 33.89, respectively.

Animals↗
PubMed2001

Genetic variation and phylogenetic analyses of the ORF5 gene of acute porcine reproductive and respiratory syndrome virus isolates.

Swine herds in the US have experienced recent outbreaks of a severe form of porcine reproductive and respiratory syndrome (designated acute or atypical PRRS) characterized by abortion and high mortality in pregnant sows. Most of the affected herds had been vaccinated with modified live-vaccines (MLVs) against PRRS. To explore the possible mechanism of the emergence of acute PRRS, the open reading frame 5 (ORF5) gene encoding the major envelope protein (GP5) of acute PRRSV isolates was characterized. The complete ORF5 gene of eight acute PRRSV isolates from herds experiencing acute PRRS outbreaks in Iowa and North Carolina was amplified and sequenced. Sequence analyses revealed that these acute PRRSV isolates shared 88-95% nucleotide and 88-96% amino acid sequence identities to each other, 87-97% nucleotide and 84-96% amino acid sequence identities with other North American PRRSV isolates and the MLVs. Most of the amino acid substitutions locate in the putative signal sequence and two short hypervariable regions at the amino terminus. The ORF5 gene sequence of the acute PRRSV isolate 98-37120-2 from a non-vaccinated swine herd in Iowa is very closely related to that of the RespPRRS MLV, with 97% nucleotide and 96% amino acid sequence identities. Phylogenetic analysis revealed that all eight acute PRRSV isolates are clustered within the North American genotype. Several minor branches that are not associated with geographic origins were also identified within the North American genotype. One acute PRRSV isolate (98-37120-2) is clustered with the RespPRRS MLV and several Danish isolates that were confirmed to be derived from the RespPRRS MLV. The ORF5 gene sequences of other seven acute isolates are more related to those of several earlier PRRSV isolates and the PrimePac MLV than to that of the RespPRRS MLV. Our results showed that the acute PRRSV isolates analyzed in this study differed from each other in ORF5 genes, although they all clustered within the North American genotype. The data from this study do not fully support the hypothesis that the emergence of acute PRRS is due to reversion of MLVs to a pathogenic phenotype, as only one of the eight acute isolates was shown to be very closely related to the RespPRRS MLV.

Acute Disease↗
PubMed1996

Comparison of the antigen distribution of two US porcine reproductive and respiratory syndrome virus isolates with that of the Lelystad virus.

One hundred 4-week-old cesarean-derived colostrum-deprived pigs were inoculated with one of two different US porcine reproductive and respiratory syndrome virus (PRRSV) isolates (VR2385, VR2431) or the European Lelystad virus to detect and compare the location and amount of virus antigen. Interstitial pneumonia, myocarditis, lymphadenopathy, and encephalitis were consistently seen in all three groups; however, disease and lesions were more severe in the VR2385 group. Immunohistochemical evaluation of formalin-fixed tissues revealed virus antigen in alveolar macrophages in lungs of 22/25, 14/25, 14/25, and 0/25 of the VR2385, VR2431, Lelystad, and control pigs, respectively. Follicular macrophages and dendritic cells in the lymph nodes of 14/25, 10/25, 10/25, and 0/25 pigs from the VR2385, VR2431, Lelystad, and control groups, respectively, stained positive for virus antigen. Similar cells in the tonsils from 25/25, 21/25, 23/25, and 0/25 pigs from the VR2385, VR2431, Lelystad, and control groups, respectively, stained positive for virus antigen. Other tissues and cells in which virus antigen was detected included macrophages and endothelial cells in the heart, macrophages, and interdigitating cells in the thymus, macrophages and dendritic cells in the spleen and Peyer's patches, and macrophages in hepatic sinusoids, renal medullary interstitium, and adrenal gland. PRRSV persisted in macrophages in the lung, tonsil, lymph node, and spleen for at least 28 days. Significantly more PRRSV antigen was detected in the lung (P < 0.01), lymph nodes (P < or = 0.05), and tonsils (P < 0.05) of the VR2385 pigs than was detected in the same tissues of the VR2431 and Lelystad pigs. The cell types in which PRRSV antigen was detected and the distribution of PRRSV antigen-positive cells within particular tissues and organs were generally similar for the different virus inoculation groups despite differences in virulence of the isolates.

Animals↗
PubMed2003

[Local recurrence of breast cancer. Isolated local recurrence located at the skin of the latissimus dorsi donor flap after breast reconstruction].

The reasons for local recurrence of breast cancer are currently unknown. Inadequate resection (not RO), undetected multifocal tumors, or absence of postoperative radiation are discussed. Secondary multifocal tumors as well as ipsilateral secondary tumors have been described as potential origins. Partial local recurrences can be considered a sign for general metastasis with an isolated local "reproduction" of the initial tumor. In our patient the observation of isolated local recurrences of the initial breast cancer, located at the skin of the latissimus dorsi donor flap, may be helpful in clarifying the ongoing controversial discussion. Our case study shows that the local recurrence occurred exactly at the initial location of the primary tumor independent of the local epidermal situation. We assume that activated vertical connections exist between the fascial lymphatic system and the dermal lymphatic plexus that "directed" the way to the body wall.

Adenocarcinoma↗
PubMed1996

Genetic variation in porcine reproductive and respiratory syndrome virus isolates in the midwestern United States.

The nucleotide sequence of a 3266 bp region encompassing open reading frames (ORFs) 2 through 7 of the porcine reproductive and respiratory syndrome virus (PRRSV) was determined for 10 isolates recovered from the midwestern United States. Pairwise comparisons showed that genetic distances between isolates ranged from 2.5% to 7.9% (mean 5.8% +/- 0.2%) whereas the Lelystad strain from Europe was, on average, 34.8% divergent from US clones. Thus, US and European PRRSV isolates represent genetically distinct clusters of the same virus. ORF 5, which encodes the envelope glycoprotein, was the most polymorphic [total nucleotide diversity (pi) = 0.097 +/- 0.007] and ORF 6, encoding the viral M protein, was the most conserved (pi = 0.038 +/- 0.003). The substantial differences in nucleotide diversity among ORFs suggests that the virus is evolving by processes other than simple accumulation of random neutral mutations. In support of this hypothesis, statistical analyses of the nucleotide sequence provided strong evidence for intragenic recombination or gene conversion in ORFs 2, 3, 4, 5 and 7, but not in ORF 6. An excess of synonymous (silent) substitutions was observed in all six ORFs, indicating an evolutionary pressure to conserve amino acid sequences. Taken together, the data indicate that despite intragenic recombination among extant PRRSV isolates, purifying selection has acted to maintain the primary structure of individual ORFs.

Animals↗
PubMed1994

Isolation and serological characterization of porcine reproductive and respiratory syndrome (PRRS) viruses from pigs with reproductive and respiratory disorders in Japan.

Three porcine reproductive and respiratory syndrome viruses (PRRSVs) were successfully isolated from stillborn piglets and fattening pigs derived from different herds affected either with an epizootic reproductive failure or a severe chronic respiratory distress in Japan. The isolates were reacted with antisera against both European (Lelystad virus) and American (strain 46448) PRRSVs in indirect immunofluorescence (IIF). However, cross immunoperoxidase monolayer assay revealed the close serological relationships between the Japanese isolates and American PRRSV, but not between the Japanese isolates and the European PRRSV. The data of preliminary serological survey with IIF technique showed a high frequency of antibody positive pigs against both Japanese isolate and the American PRRSV, whereas a low frequency of antibody positive pigs against the European PRRSV, on herds which have had clinical episodes of PRRS-like diseases. These results indicate that PRRSVs are prevalent in Japan, and suggest that the antigenicities of the prevalent PRRSVs are more closely related to those of the American than the European PRRSVs.

Animals↗
PubMed1984

[Morphology and problems of the morphogenesis of cytomegalic sialadenitis].

Two varieties of nodular sialadenitis, acute nodular (15%) and chronic with exacerbation of the nodular type (19%), were distinguished basing on analysis of the material from 880 children dead from cytomegaly. Parotid glands were examined electron-microscopically in 2 cass of generalized and 6 cases of localized cytomegaloviral infection. Basing on the histoenzymochemical data published earlier, the etiological role of cytomegalovirus in the development of nodular infiltrates was substantiated and the functional activity of cytomegalic cells in virus reproduction and isolation shown. Diagnostic importance of nodular infiltrates was stressed. Morphological manifestations of cytomegalovirus reproduction were described.

Cell Transformation, Viral↗
PubMed1981

Isolation of Pasteurella ureae from reproductive tracts of congenic mice.

Infertility noted in two congenic strains of mice was associated with abortion, metritis, and stillbirths. Pasteurella pneumotropica was recovered from affected uterine tracts of ATL mice. Tetracycline treatment for 6 weeks was instituted in an attempt to minimize Pasteurella-associated infertility. Nine ATL mice and 12 ATH mice were examined 4 and 12 months after cessation of tetracycline therapy, respectively. Histopathologically, all animals appeared normal. P. pneumotropica was recovered from six of nine prepuces of both ATH and ATL mice; similarly, two of three vaginal culture of the ATL mice yielded this bacterium. Pasteurella ureae was isolated from three of eight reproductive tracts of the female ATH mice, Negative indole and ornithine decarboxylase reactions and positive acid production for mannitol differentiated P. ureae from P. pneumotropica. This report represents the first confirmed isolation of P. urea from animals. Biochemical profiles of atypical strains of P. pneumotropica isolated from animals are discussed and compared with those of P. ureae. The need for careful biochemical characterization of all Pasteurella isolated from rodents is emphasized.

Animals↗
PubMed1998

Differentiation between porcine reproductive and respiratory syndrome virus isolates by restriction fragment length polymorphism of their ORFs 6 and 7 genes.

Three distinct antigenic profiles were identified by comparing the reactivities of 15 Canadian field isolates, the attenuated U.S. vaccine (Ingelvac MLV) strain and 2 European reference strains (Lelystad and Weybridge) of the porcine reproductive and respiratory syndrome virus (PRRSV) by indirect immunofluorescence with a set of 4 monoclonal antibodies to the nucleocapsid (N) protein and 2 other to the matrix (M) protein. In the present study, 9 Canadian isolates for which the sequences were determined appeared closely related to 2 U.S. reference strains (ATCC VR-2332 and ATCC VR-2385) with amino acid identities varying between 90 to 98% for the M and N proteins; substitutions in the nucleotide sequences were distributed randomly throughout the ORFs 6 and 7 genes, and most were 3rd base silent mutations. In comparison, more than 30% divergence was demonstrated with the Lelystad virus. Furthermore, differentiation between North American and European isolates, and between field isolates and the MLV strain could be achieved by cutting PCR-amplified products encompassing both ORFs 6 and 7 genes with 4 restriction endonucleases. When taken individually, BsaJI and AluI were the more appropriate restriction enzymes for distinguishing the vaccine strain from field isolates. The results obtained suggest that the restriction fragment length polymorphism of the genomic region covering the ORFs 6 and 7 genes may be a valuable tool to differentiate among PRRSV isolates.

Animals↗
PubMed1999

Factors affecting reproductive performance in captive Mallard ducks.

Propagation of wild-strain Mallards (Anas platyrhynchos) in captivity is hindered by low egg fertility. Therefore, we studied the effects of captivity, age, mate choice and isolation on reproductive parameters of Mallards. Captive drakes had smaller immature testes than free-flying Mallard drakes. Captive yearling ducks weighed less than adult ducks at the beginning of the breeding season, but no differences were found between their initial clutch size, egg volume or number of clutches laid. Yearling pairs had lower egg fertility (7%) than adult pairs (80%). Egg fertility was higher (51 vs 21%) in self-chosen pairs than in randomly assigned pairs. Isolation of ducks, however did not influence egg production.

Aging↗
PubMed2006

Income breeding allows an aquatic snake Seminatrix pygaea to reproduce normally following prolonged drought-induced aestivation.

1. Capital breeding is an ideal reproductive strategy for many ectotherms because it provides a disassociation between feeding and reproduction, a necessary requirement for animals that become anorexic during pregnancy. Among ectotherms, some viviparous snakes (e.g. Viperidae) exemplify the capital breeding strategy because many species (i) do not feed during pregnancy due to behavioural conflicts between reproduction and foraging, and (ii) take more than one season to accumulate sufficient energetic stores for reproduction. 2. Isolated wetlands often exhibit extreme annual fluctuations in environmental conditions with prolonged droughts periodically leaving wetlands completely dry and devoid of prey. Following droughts, however, wetlands can be extremely productive, rendering prey resources virtually unlimited for some species. 3. This study examines drought survival strategy and reproductive ecology of a small aquatic snake Seminatrix pygaea (Cope) in an isolated wetland. Seminatrix pygaea are atypical from most sympatric snake species in that (i) their small body size, reliance on aquatic prey, and high rates of evaporative water loss make them ill-suited to overland movement, and (ii) they may not be subject to costs typically associated with feeding during pregnancy. 4. We hypothesized that S. pygaea would survive periodic multiyear droughts by aestivating within the dried wetland, a survival strategy heretofore undocumented in snakes. Further, we hypothesized that if S. pygaea rely on 'typical' snake reproductive strategies of 'adaptive anorexia' and capital breeding, reproductive output would be reduced in the first wet year following drought. 5. By encircling a 10-ha wetland with a continuous drift fence before it refilled we were able to demonstrate that S. pygaea were present within the dried wetland prior to the onset of spring rains that refilled the wetland in 2003. Our results suggest that S. pygaea are capable of surviving multiyear droughts by aestivating within the dried wetland. 6. Despite having presumably depleted energy reserves during the drought, S. pygaea reproduced with the same frequency and fecundity during the first season following refilling of the wetland as in pre-drought years. 7. The ability of S. pygaea to rebound rapidly from the stresses of prolonged drought is due in part to their reproductive ecology. Seminatrix pygaea readily feed throughout pregnancy and consequently can rapidly translate high prey abundances into reproductive output through income breeding.

Animals↗
PubMed1999

Study of preantral follicle population in situ and after mechanical isolation from caprine ovaries at different reproductive stages.

The purposes of this study were to estimate the population of caprine preantral follicles, and to evaluate quantitatively and qualitatively the efficiency of a specific mechanical method for the isolation of preantral follicles from mixed breed goats at different reproductive stages. On average, 37,646+/-4277 preantral follicles were present in goat ovaries, and 13,631+/-2399 preantral follicles were obtained after isolation. The number of preantral follicles isolated or in situ was not significantly affected by the reproductive stage. The mean recovery rate per ovary ([number of isolated follicles/number of in situ follicles] x 100) of isolated follicles was 36.2%. The distribution of follicles in situ was 67.8% primordial, 25.8% primary and 6.4% secondary; the respective distribution after isolation was 93.8%, 5.2% and 1.0%. In this study, many polyovular follicles were also observed, mainly in prepubertal goat ovaries. Histological analysis showed that few preantral follicles were atretic in situ (4.83%+/-0.35) or after the isolation procedure (4.67%+/-0.65) in the three reproductive stages. The percentage of atretic follicles was not affected either by the mechanical method or by the reproductive stage. It is concluded that a large number of preantral follicles can be successfully isolated mechanically, with a high recovery rate and a low rate of follicular atresia, irrespective of the reproductive stage of the caprine female.

Age Factors↗
PubMed1998

Clinical consequences of exposing pregnant gilts to strains of porcine reproductive and respiratory syndrome (PRRS) virus isolated from field cases of "atypical" PRRS.

OBJECTIVE: To determine clinical consequences of exposing pregnant gilts to strains of porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) isolated from field cases of "atypical" or "acute" PRRS in vaccinated herds. ANIMALS: 20 pregnant gilts and their pigs and fetuses. PROCEDURE: 8 pregnant gilts (principals: 4 groups [2 gilts/group]) were exposed oronasally at or about 45 days of gestation to 1 of 4 strains of PRRSV and necropsied 6 weeks later. Nonexposed controls (2 additional pregnant gilts) were kept under otherwise similar conditions. The experiment was repeated, except that principals were exposed at or approximately 90 days of gestation and allowed to farrow. Clinical observations were made at least twice daily, and samples and specimens from gilts and their fetuses and pigs were tested for PRRSV and homologous antibody. RESULTS: Exposure of pregnant gilts to PRRSV at or approximately 45 days' gestation resulted in low prevalence of transplacental infection and fetal death. Exposure of pregnant gilts to PRRSV at or approximately 90 days' gestation resulted in higher prevalence of transplacental infection and fetal death. Moreover, 1 gilt aborted and many liveborn pigs of other litters were weak and unthrifty. Clinical signs of disease and reproductive failure were especially severe for a field strain of PRRSV isolated from an epizootic that fit the strictest definition of atypical PRRS. Controls remained clinically normal and free of PRRSV. CONCLUSION AND CLINICAL RELEVANCE: Some strains of PRRSV now circulating in US swine herds are more virulent than those encountered in the past. Clinical PRRS in vaccinated herds suggests need for a new generation of vaccines.

Abortion, Veterinary↗
PubMed1981

Combination antimicrobial therapy for serious gynecological and obstetrical infections--obsolete?

The normal flora of the lower reproductive tract are the pathogens isolated from upper reproductive tract infections after cesarean section and hysterectomy and from intraperitoneal cultures taken from women treated for acute pelvic inflammatory disease. Because these infections are usually polymicrobial, traditional antimicrobial therapy has included various combinations of agents. During the last six years we used five different antimicrobial combinations to treat more than 600 women with one of the above infections. From 15% to 50% of cases required the addition of a third (anaerobic-specific) antimicrobial. Original therapy with clindamycin/gentamicin was unsuccessful in only 5% of the infections treated. Single-agent therapy with third-generation cephalosporins avoids most problems associated with combination therapy. Over the past 18 months we treated 237 women with cefotaxime; 94% required no further therapy.

Anaerobiosis↗
PubMedDate not supplied

Antigenic variability of porcine reproductive and respiratory syndrome (PRRS) virus isolates. Influence of virus passage in pig.

In order to study the antigenic variability of porcine reproductive and respiratory syndrome virus (PRRSV), 18 European and Canadian field isolates were analysed with a panel of 15 monoclonal antibodies (MAbs) raised against five different European and one Canadian PRRSV isolates. The antigenic pattern of these isolates was used to infer their phylogenetic relationships. Isolates which had the same reactivity pattern were gathered together so that five antigenic profiles were analysed. The pairwise distances between these groups were defined based on antigenic pattern differences. Two main antigenic groups were obtained, discriminating between the European and the Canadian populations, as illustrated by the great distance observed between European and Canadian isolates (D = 0.5619 +/- 0.0625) compared to the distance between European isolates (D = 0.1524 +/- 0.0735). The distance matrix allowed also the construction of a tree diagram. Bootstrap analysis was performed to test the confidence in the branching. The tree diagram confirmed the distinction between the European and the Canadian PRRSV populations. Antigenic variability between an isolate and its progeny recovered after one or two passages in vivo was examined on six isolates. It was restricted to the GP3 protein of the virus.

Animals↗