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PubMed1994

Molecular cloning and nucleotide sequencing of the 3'-terminal genomic RNA of the porcine reproductive and respiratory syndrome virus.

The genomic RNA of a porcine reproductive and respiratory syndrome virus (PRRSV) isolate from the U.S.A., VR 2385 (ATCC), was copied into cDNA after priming with oligo(dT) and cloned into phage lambda. The cDNA clones representing the 3'-terminal genomic RNA of the virus were isolated and sequenced. The genome is a positive-stranded, polyadenylated RNA with an estimated size of 15 kb. Analysis of the resulting sequence identified three complete open reading frames (ORFs) with the potential to encode polypeptides with predicted M(r)s of 22.2K (ORF 5), 19.1K (ORF 6) and 13.6K (ORF 7). ORF 7, which is closest to the 3' end, is predicted to encode a highly basic nucleocapsid protein displaying 58% amino acid identity to the corresponding protein of the Lelystad virus (LV), a European PRRSV isolate. ORFs 6 and 5, preceding ORF 7, are each predicted to encode proteins containing several hydrophobic domains that are thought to be membrane-associated. The VR 2385 ORF 6 protein is the most conserved structural protein. It has 78% amino acid identity to the equivalent LV protein, and ORF 5 shares only 54% of its amino acid sequence. Northern blot analysis revealed a 3'-coterminal nested set of six subgenomic RNAs in VR 2385 virus-infected CRL 11171 cells. Our results indicate that VR 2385, like LV, is a member of the newly proposed arterivirus group. However, the striking genetic variation and the difference in pathogenicity between LV and VR 2385 suggest that the viruses causing PRRS in the U.S.A. and Europe are highly variable and they may represent different genotypes.

Amino Acid Sequence↗
PubMedDate not supplied

Conjugation-specific genes in the ciliate Euplotes crassus: gene expression from the old macronucleus.

Following mating or conjugation, the hypotrichous ciliate Euplotes crassus undergoes a massive genome reorganization process. While the nature of the rearrangement events has been well studied, little is known concerning proteins that carry out such processes. As a means of identifying such proteins, differential screening of a developmental cDNA library, as well as construction of a cDNA subtraction library, was used to isolate genes expressed only during sexual reproduction. Five different conjugation-specific genes have been identified that are maximally expressed early in conjugation, during the period of micronuclear meiosis, which is just prior to macronuclear development and the DNA rearrangement process. All five genes are retained in the mature macronucleus. Micronuclear, macronuclear, and cDNA clones of one gene (conZA7) have been sequenced, and the results indicate that the gene encodes a putative DNA binding protein. In addition, the presence of an internal eliminated sequence in the micronuclear copy of the conZA7 gene indicates that this conjugation-specific gene is transcribed from the old macronucleus.

Amino Acid Sequence↗
PubMed1986

Investigation of purulent vaginal discharge in cows, with particular reference to Haemophilus somnus.

The relationship between vaginal discharge, site of inflammation in the reproductive tract and species of bacteria isolated was examined in cows that were either normal, subfertile, or had had an abnormal parturition. Microbiological and cytological examinations were performed on uterine swabs from 221 cows and cervical swabs from 107 cows. There were 167 subferitle cows, 144 of which had failed to conceive to 3 or more services (FTC) and 23 of which had not had a visible oestrus since calving (NVO). Metritis was diagnosed in 22% of the FTC cows, 48% of the NVO cows, and 12% of the cows with abnormal parturition. Fifty percent of cows with metritis had purulent vaginal discharge. However, only 25% of a total of 91 cows with vaginal discharge had metritis; the remainder had cervicitis. Fifty-four percent of subfertile cows had cervicitis without metritis. Mixed infection with aerobes and anaerobes was significantly correlated with uterine inflammation and was found in 20% of cows with metritis. The rate of isolation of H.somnus from inflamed uteruses (22%) and inflamed cervixes (39%) was significantly greater than the isolation rate from normal uteruses (8%) and cervixes (10%). No bacteria were isolated from 50% of inflamed uteruses and 37% of inflamed cervixes. The number of subfertile cows with or without metritis that subsequently conceived was not significantly different.

Animals↗
PubMed1979

Induction of Listeria monocytogenes infection by the consumption of ponderosa pine needles.

An infectious microorganism, identified as Listeria monocytogenes, has been isolated from the bloodstream of pregnant mice fed a diet containing Pinus ponderosa needles. When the isolate was injected into pregnant mice, reproductive dysfunction and other changes, including speckled livers, spleen atrophy, and hemorrhagic intestines, appeared to mimic the signs of the disease in pregnant mice fed pine needles. Moreover, these pathological changes are similar to those observed in cattle and other mammals experiencing abortions or toxemia, or both, attributed to the ingestion of P. ponderosa needles, suggesting that L. monocytogenes may be a part of the etiology of "pine needle abortion."

Abortion, Veterinary↗
PubMed1995

Persistence of porcine reproductive and respiratory syndrome virus in serum and semen of adult boars.

Four seronegative adult boars were intranasally inoculated with porcine reproductive and respiratory syndrome virus (PRRSV) isolate VR-2332. Serum and semen were collected 2-3 times weekly for over 100 days postinoculation (DPI). Serum samples were assayed for PRRSV by virus isolation (VI) and a polymerase chain reaction (PCR) and screened for antibodies to PRRSV using the indirect fluorescent antibody (IFA) and virus neutralization (VN) tests. Semen was assayed for PRRSV RNA by PCR. Virus and viral RNA was detected in the serum of all boars within 1 DPI by Vi and/or PCR. However, VI results indicated that viremia was transient and occurred from 1 to 9 DPI. Viral RNA was detected in serum from 1 to 31 DPI. In the acute stage of the infection, PRRSV RNA was detected in serum by PCR prior to the presence of viral RNA in semen. The PRRSV RNA was detected in semen as early as 3 DPI and persisted for 25 DPI in 2 of the boars and 56 and 92 DPI in the remaining 2 boars. Detection of PRRSV RNA in semen occurred 2-8 and 28-35 days prior to the detection of antibodies by IFA and VN, respectively. PRRSV was isolated from the bulbourethral gland of the boar that shed viral RNA in semen for 92 DPI. These results suggest that PRRSV RNA can be detected by PCR in boar serum and semen, and may persist for variable periods of time. Viremia and the serologic status of the boar are not adequate indicators of when PRRSV or PRRSV RNA is being shed in the semen. Preliminary findings also indicated that neither shipping stress nor reinoculation with homologous PRRSV resulted in viremia or viral RNA shedding in semen.

Animals↗
PubMed2000

[The microbial findings in leukorrhea].

During two periods /1984/86 and 1998/01.00/ were studied microbial findings from vaginal specimens of women in reproductive age /16-48/. Candida spp. was isolated most frequently /17.3% to 21.5%/. BV was increased from 4.6% to 13.1%. T. vaginalis was decreased from 8.7% to 1.8%, probably as a consequence of metronidazole appliance.

Adolescent↗
PubMed1992

[The formation of host-parasite relationships in an experimental mixed pathology of opisthorchiasis-tuberculosis as dependent on the phase of the Opisthorchis infestation].

Superinfection of animals having Opisthorchis invasion with Mycobacterium tuberculosis leads to destabilization of host-parasite relationships in opisthorchiasis and formation of new host-parasitocenotic interrelations whose manifestation depends on the phase of the invasive process. At the acute invasive phase of mixed pathology (2 weeks) the activity of the host's immune system increases, while the biological activity of helminths and the number of M. tuberculosis colonies decrease. And on the contrary, at the subacute phase of mixed pathology (2.5 months) the activity of the host's immune system decreases, while the reproductive activity of helminths and the isolation rate of M. tuberculosis from pulmonary tissue increases in comparison with the groups of animals with monoinvasion and monoinfection.

Acute Disease↗
PubMed2003

Sequence diversity of mating-type genes in Phaeosphaeria avenaria.

Phaeosphaeria avenaria, one of the causal agents of stagonospora leaf blotch diseases in cereals, is composed of two subspecies, P. avenaria f. sp. triticea (Pat) and P. avenaria f. sp. avenaria (Paa). The Pat subspecies was grouped into Pat1-Pat3, based on restriction fragment length polymorphism (RFLP) and ribosomal DNA (rDNA) internal transcribed spacer (ITS) sequences in previous studies. Mating-type genes and their potential use in phylogeny and molecular classification were studied by DNA hybridization and PCR amplification. The majority of Pat1 isolates reported to be homothallic and producing sexual reproduction structures on cultural media had only the MAT1-1 gene. Minor sequence variations were found in the conserved region of MAT1-1 gene in Pat1 isolates. However, both mating-type genes, MAT1-1 and MAT1-2, were identified in P. avenaria isolates represented by ATCC12277 from oats (Paa) and the Pat2 isolates from foxtail barley ( Hordeum jubatum L.). Cluster analyses based on mating-type gene conserved regions revealed that cereal Phaeosphaeria is not phylogenetically closely related to other ascomycetes, including Mycosphaerella graminicola (anamorph Septoria tritici). The sequence diversity of mating-type genes in Pat and Paa supports our previous phylogenetic relationship and molecular classification based on RFLP fingerprinting and rDNA ITS sequences.

Ascomycota↗
PubMed1975

T-strain mucoplasmas and reproductive failure in monkeys.

T-strain mycoplasmas were isolated from the genital tracts of 3 genera of female monkeys: the talapoins, Miopithecas talapoin, the patas, Erythrocebus patas, and the macaques, Macaca fascicularis. The talapoins have not reproduced successfully for the past 2 yr. Fertility in the patas has also been reduced, with a high incidence of stillbirths and spontaneous abortions. The macaques were not permitted to breed.

Abortion, Veterinary↗
PubMed1982

Biological characteristics and host influence on a geographical isolate of Trichinella (wolverine: 55 degrees 00'N, 100 degrees 00'W, 1979).

Biological characteristics of Trichinella isolate (wolverine: 55 degrees 00'N, 100 degrees 00'W, 1979) were established in Crl: COBS CFW (SW) mice. Comparison of the wolverine isolate's biological characteristics with another Trichinella isolate (polar bear), both from closely related geographic areas, revealed there were stable and reproducible genetic differences between isolates. Differences were most pronounced for degree of infections causing a 50% mortality of mice, larval production by females in vitro, reproductive capacity indices, and survival of muscle larvae. Species and strain of host altered characteristics of the wolverine isolate such as worm position, sex ratios, % recovery, larval production by females in vitro, and reproductive capacity indices. Genetic differences between isolates of Trichinella and interaction with host genetics raises interesting questions on Trichinella speciation. Differences reported here, are best interpreted for the time being at least, as part of the normal biological variability of the species Trichinella spiralis.

Animals↗
PubMed2006

Molecular and functional characterization of a novel gonadotropin-releasing-hormone receptor isolated from the common octopus (Octopus vulgaris).

GnRH (gonadotropin-releasing hormone) plays a pivotal role in the regulation of reproduction in vertebrates through interaction with a specific receptor. Previously, we isolated a GnRH homologue, oct-GnRH, from the common octopus (Octopus vulgaris). In the present study, we have identified a GnRH receptor (oct-GnRHR) specific for oct-GnRH from Octopus brain. Oct-GnRHR includes domains and motifs typical of vertebrate GnRH receptors. The intron-inserted positions are conserved between oct-GnRHR and the chordate GnRHR genes. The oct-GnRHR expressed in Xenopus (South African clawed frog) oocytes was responsive to oct-GnRH, but not to any other HPLC fractions of the Octopus brain extract. These results show that oct-GnRHR is an authentic receptor for oct-GnRH. Southern blotting of reverse-transcription PCR products revealed that the oct-GnRHR mRNA was widely distributed in the central and peripheral nervous systems and in several peripheral tissues. In situ hybridization showed that oct-GnRHR mRNA was expressed in some regions involved in autonomic functions, feeding, memory and movement. Oct-GnRH was shown to induce steroidogenesis of testosterone, progesterone and 17beta-oestradiol in Octopus ovary and testis, where oct-GnRHR was abundantly expressed. These results suggest that oct-GnRH, like its vertebrate counterparts, acts as a multifunctional neurotransmitter, neuromodulator and hormone-like factor, both in Octopus central nervous system and peripheral tissues, and that both structure and functions of the GnRH family are, at least partially, evolutionarily conserved between octopuses and chordates.

Amino Acid Sequence↗
PubMed1999

Epidemiologic, pathogenic and molecular analysis of recent encephalomyocarditis outbreaks in Belgium.

In 1991 EMCV was isolated for the first time in Belgium from the offspring of a sow with reproductive failure. From August 1995 until December 1996, EMCV was diagnosed in 154 Belgian pig holdings in association with myocardial failure and sudden death in fatteners and suckling piglets or with reproductive failure in sows. To clarify some epidemiological aspects 3 EMCV isolates characteristic for the different clinical pictures and outbreaks were studied. Field observations and animal experiments indicated that the pathogenicity induced by each isolate is specific for one age category and that the spread of the virus is limited. The presented data also suggest that rodents may play a role in the transmission of EMCV but that pig-to-pig transmission is at least as important. Molecular analysis of two separate regions on the genomes of the respective EMCV isolates showed that the 1995-96 EMCV epizootic in Belgium was due to a new virus introduction. Furthermore, the VP1 coding gene is proposed as a marker of virulence.

Amino Acid Sequence↗
PubMed1984

Susceptibility profiles of potential aerobic and anaerobic pathogens isolated from hysterectomy patients.

A total of 1140 aerobic and anaerobic isolates were recovered from cultures of specimens from the reproductive tracts of 435 uninfected patients who underwent elective hysterectomy. Standard minimum inhibitory concentration susceptibility studies were performed on these isolates to 13 newer penicillins, cephalosporins, and some traditional antimicrobial agents. These data were generated to evaluate the in vitro efficacy of these antibiotics for potential use in prophylaxis or as a single agent for treatment of polymicrobial infections of the female pelvis. The minimum inhibitory concentration data for each antibiotic against 16 genera of aerobic and nine genera of anaerobic bacteria were determined and were used to compare the in vitro antimicrobial activity of newer antibiotics to that of the more traditional antibiotics. Of the antimicrobial agents tested, piperacillin had the highest in vitro activity against these isolates of any antibiotic tested.

Anti-Bacterial Agents↗
PubMed1986

Acid phosphatases in germinal and somatic cells of the testes.

Four forms of acid phosphatase have been found in the testicular tissue of many mammalian species, but their exact cellular site has remained obscure. In this work, acid phosphatases have been studied in different reproductive organs of the male rat, in somatic cell lines derived by cloning from both rat and mouse testes, in primary cultures of rat Sertoli cells, and in isolated spermatogenic cells of the mouse. Among the reproductive organs, preputial glands show the highest specific activities with p-nitrophenyl phosphate as substrate, followed by the testicular tissue and the different regions of the epididymis. By contrast to that in other tissues, testicular activity with p-nitrophenyl phosphate is not influenced by tartrate and is activated markedly by cobalt (Co2+). Among the somatic cell lines, the highest hydrolysis rates are obtained with naphthyl substrates in the epithelial (TR-1) and myoid (TR-M) cell lines and marginally lower rates in the Leydig (TM3) and Sertoli (TM4) cell lines. With thymolphthalein phosphate, the latter two cell lines show very low activity. These activities are not influenced by different hormones and growth factors in the culture medium. The most marked Co2+-activated reaction with p-nitrophenyl phosphate is found in advanced stages of germinal cells and residual bodies. Primary cultures of Sertoli cells, prepared from rats 10 to 30 days of age, show a slight decrease in acid phosphatase levels; however, the activities are not influenced markedly by addition of follicle-stimulating hormone (FSH) and/or testosterone to the culture medium.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid Phosphatase↗
PubMed1999

Isolated pleural effusion in severe ovarian hyperstimulation: A case report.

Assisted reproductive technology programs use controlled ovarian hyperstimulation to maximize pregnancy rates. Severe ovarian hyperstimulation syndrome is a well-known risk. Pleural effusion often accompanies severe ovarian hyperstimulation syndrome. We describe 2 cases of isolated hydrothorax without concomitant ascites and review the literature of this rare finding.

Abortion, Spontaneous↗
PubMed1997

Survival of Trypanosoma musculi in the kidneys of chronically infected mice: kidney form reproduction and immunological reactions.

Trypanosoma musculi are detected in the blood of the mouse host within 3-5 days after infection. Peak parasitemia is reached within 10 days and parasites persist at a plateau level for 2-3 wk. There are no intracellular stages and the flagellates are eliminated from the peripheral blood within 4 wk. However, trypanosomes persist in the vasa recta of the kidneys and may be present for the life of the host. Infection provides lifelong resistance to reinfection. Kidney forms (KF) of T. musculi were isolated and studied to define their morphology, reproductive activity, and serological reactivity. Dividing epimastigotes and trypomastigote stages were present in kidneys. Multinucleate and rosette forms were common. Measurements of the coefficient of variation of the KF confirmed that the trypanosomes were actively reproducing. Direct immunofluorescence reactions with rabbit antimouse IgG + IgA + IgM detected antibodies on bloodstream forms (BSF) but not on the KF. However, indirect immunofluorescence tests using antisera collected from mice during early and late phases of the infections showed that KF were capable of reacting with antibodies. The reproductive activity displayed by the KF appears to be responsible for the continuous replacement of the trypanosomes that are killed by the immune responses of the resistant mice when they leave the vasa recta. Solute concentrations of the blood within the vasa recta appear to prevent antibodies from complexing with surface antigens of the parasites. These capillaries provide nutrients that allow the trypanosomes to reproduce and persist unaffected by the host immune responses.

Animals↗
PubMed2000

Preliminary findings of Salmonella spp. in captive green iguanas (Iguana iguana) and their environment.

Captive reptiles are routinely identified as reservoirs of Salmonella spp. and reports of reptile-associated salmonellosis are increasing. Unfortunately, little is known about the epidemiology of Salmonella spp. and green iguanas. We did a limited survey of a green-iguana farm in El Salvador to identify sources of Salmonella spp. in green iguanas and their environment. A limited number of samples for microbiological culture were collected from iguanas (adult, hatchling, and embryos) and their environment (food, water, soil, shelter, insects, and wild-caught lizards). Salmonella spp. was isolated from the intestine of both adult (3/20) and hatchling iguanas (8/20). There was no evidence of Salmonella spp. in the reproductive tracts of female iguanas (0/10). Salmonella spp. was isolated from the surface of 40% (7/16) of the egg surfaces tested. Salmonella spp. was not identified from the externalized yolk-sac of the iguana embryos tested. Soil samples from a breeding pen and a nest were both positive for Salmonella spp. Eight different Salmonella spp. serotypes were identified in this survey. These results suggest that horizontal transmission of Salmonella spp. is a potential source of exposure to hatchling iguanas at this facility.

Animals↗