Effects of the administration of ketoprofen at parturition on the milk yield and fertility of Holstein-Friesian cattle.

A total of 220 cows and heifers were treated with 3 mg/kg ketoprofen immediately after calving and 24 hours later, and 227 were left untreated as controls. The treated animals tended to have a lower incidence of retained fetal membranes and were 1.7 times less likely to develop the condition than the untreated animals. The treatment did not affect the incidence of milk fever, the endometritis score or the presence of a corpus luteum by 20 to 25 days after calving, and did not affect the cows' early lactation milk yield. There was no particular level of dystocia or periparturient conditions other than retained fetal membranes for which there might be a reproductive or productive advantage following the use of ketoprofen. The intervals from calving to first insemination or to pregnancy, the numbers of inseminations per pregnancy and the pregnancy rate to first insemination were also unaffected by ketoprofen treatment.

Diagnostic exercise: papulovesicular dermatitis in rhesus macaques (Macaca mulatta).

Eleven rhesus monkeys developed multifocal erythematous and a vesicular rash. Most recovered spontaneously, but a 21-year-old female became moribund and was euthanized. Findings were of vesicular dermatitis and widespread multifocal hemorrhagic necrosis of the lungs and other viscera, with intralesional intranuclear inclusions. Simian varicella virus was identified as the cause by polymerase chain reaction analysis and serologic testing.

Primary mouse dermal fibroblast cell cultures as an in vitro model system for the differential pathogenicity of cross-species herpesvirus papio 2 infections.

Infection of mice with herpesvirus papio 2 (HVP2) parallels zoonotic monkey B virus infections. A major benefit of the HVP2/mouse model is the existence of two HVP2 subtypes: HVP2nv rapidly invades and destroys the CNS while HVP2ap produces no clinical signs and mild histopathological lesions. However, in the natural baboon host, no difference in pathogenicity of HVP2 subtypes is evident. Primary dermal fibroblast cells were evaluated as a model system for defining virus-host interactions that influence the outcome of a cross-species infection. No differences in plaque formation or virus replication were observed between HVP2 subtypes in primary baboon dermal fibroblast cultures. In contrast, when primary mouse dermal fibroblasts (PMDF) were infected, HVP2nv replicated to higher titers and was more efficient at shutting down host-cell protein synthesis compared to HVP2ap. HVP2ap-infected PMDF cells produced more IFN-beta compared to HVP2nv, and IFN-beta pretreatment of PMDF cultures inhibited HVP2ap replication but did not affect HVP2nv. The differential pathogenicity of HVP2 subtypes in mice and the lack of such differences in the natural baboon host are recapitulated in the primary dermal fibroblast cell culture system. This model may prove useful in examining early, local, host responses that influence the outcome of cross-species infections.

Effects of three types of trace element supplementation on the fertility of three commercial dairy herds.

The effects on the fertility of three commercial dairy herds of three types of copper- and selenium-containing mineral supplements was investigated. As the cows on each farm were dried off they were allocated to one of three treatment groups, and treated with either subcutaneous injections of copper and selenium, or two matrix intraruminal trace element boluses, or two glass intraruminal trace element boluses. When the data from the 406 cows on the three farms were combined, there was a significant difference between the conception rates of the three groups (P < 0.001). The cows treated with the glass boluses conceived at a rate 1.8 times greater than those treated by injection (P < 0.001), and at a rate 1.5 times greater than those treated with matrix boluses (P = 0.002). These differences were associated with a significantly higher likelihood of service resulting in a conception in the group treated with glass boluses than in the group treated by injection (P = 0.004). After adjusting for time from calving, time from treatment, time of year and farm, there was a significant (P = 0.012) difference in glutathione peroxidase activities between the treatments, with the group treated by injection having a significantly lower activity than the groups treated with boluses.

Sequence and genetic arrangement of the UL region of the monkey B virus (Cercopithecine herpesvirus 1) genome and comparison with the UL region of other primate herpesviruses.

The complete DNA sequence of the unique long (U(L)) region of monkey B virus (BV) was determined. Based on sequence homology and the presence of transcriptional control element motifs, homologues of every open reading frame present in the U(L) region of the Human herpesvirus 1 (herpes simplex virus 1, HSV-1) and Human herpesvirus 2 (herpes simplex virus 2, HSV-2) genomes were identified in BV. The BV genes are arranged in the same order and orientation as in HSV. These results demonstrate that the BV U(L) region is entirely co-linear with that of HSV-1 and HSV-2.

Construction and in vivo detection of an enhanced green fluorescent protein-expressing strain of Saimiriine herpesvirus 1 (SaHV-1).

Saimiriine herpesvirus 1 (SaHV-1) is an alpha-herpesvirus of squirrel monkeys used in mice to study neural pathogenesis of herpesviruses. To trace dissemination of virus from a peripheral site of inoculation to the central nervous system tissues, a recombinant strain of SaHV-1 expressing enhanced green fluorescent protein (GFP) was constructed by site-specific insertion of a GFP expression cassette into a transcriptionally null point in the SaHV-1 genome. PCR and Southern blot confirmed insertion of a single GFP expression cassette into the target site of the SaHV-1 genome. The recombinant virus was shown to produce strong fluorescence in the cytoplasm of infected cells in vitro. Growth kinetic experiments demonstrated no differences between recombinant and wild type SaHV-1 in producing infectious progeny virions. The recombinant virus was comparable to wild type SaHV-1 in development of clinical disease, microscopic lesions and induction of an antibody response in mice following intramuscular inoculation. Using confocal microscopy, GFP expression was easily observed in formalin fixed, paraffin-embedded tissues of mice infected with the recombinant SaHV-1. This simple specimen processing technique preserves tissue morphology and allows detection of viral replication within various tissues of experimentally infected animals.

Simian foamy virus infections in a baboon breeding colony.

The prevalence, transmission, and variation of simian foamy viruses (SFVs) in baboons was investigated. Over 95% of adult baboons in the breeding colony as well as recently imported adult animals had high titers of anti-SFV serum IgG. Maternal antibody was detectable in infants' serum up to 6 months of age. Approximately 30% of infants in breeding harems experienced SFV infections by 1 year of age. Shedding of SFV in oral secretions was common, with 13% of samples from normal adult animals and 35% from immunosuppressed animals containing infectious SFV. SFV was isolated from three baboon subspecies (olive, yellow, and chacma baboons) and sequences from both the pol and the LTR regions of the provirus were amplified by PCR and sequenced. Phylogenetic analysis indicated that all baboon isolates formed a single lineage distinct from SFVs of other African monkey species. Within the baboon SFV lineage, two distinct clades were apparent, which consisted of isolates from yellow and olive baboons and isolates from chacma baboons. Competition ELISAs indicated that, while SFV isolates of these two groups were very closely related, antigenic differences do exist between them. SFV isolates from a drill and a mandrill were distinct from baboon SFV isolates, both genetically and antigenically.

Retrospective analysis of an outbreak of B virus infection in a colony of DeBrazza's monkeys (Cercopithecus neglectus).

In 1981, an outbreak of herpetic disease developed in a colony of DeBrazza's monkeys (Cercopithecus neglectus). In seven of eight infected animals, clinical signs of infection included vesicular and ulcerative lesions on the lips, tongue, and/or palate. Histologic examination of lesions revealed intranuclear inclusion bodies, and electron microscopy revealed nucleocapsids and virions with typical herpesvirus morphology. Although a virus was isolated that appeared similar to monkey B virus, techniques available at the time did not allow precise identification of the virus. Analysis of serum from one surviving monkey collected 12 years after the outbreak revealed a pattern of reactivity characteristic of B virus-positive serum on the basis of results of ELISA and western immunoblot analysis. Polymerase chain reaction analysis of archived paraffin-embedded tissue specimens and molecular analysis of the one viral isolate obtained from a DeBrazza's monkey indicated that the virus responsible for the outbreak was a new genotype of B virus. Testing of sera from lion-tailed macaques (Macaca silenus) housed in an adjacent cage at the same zoo indicated that these animals harbored this virus and, thus, were the likely source of the virus that infected the DeBrazza's monkeys. This study documents usefulness of archiving samples from disease outbreaks for later analysis. In addition, this incident underscores the importance of considering herpes B virus infection when outbreaks of disease having characteristics of herpetic infections develop in nonhuman primates kept at institutions that also house macaques.

Effects of priming positive and negative outcomes on drinking responses.

The purpose of this investigation was to study the effects of priming positive and negative expectancy outcomes on the drinking responses of college students. Men and women (N = 64) were randomly assigned to 1 of 3 priming conditions: a positive expectancy outcome condition, a negative expectancy outcome condition, and a neutral (control) condition. Participants were exposed to a series of semantic primes corresponding to their condition and then asked to complete a beer taste-rating task. Planned comparisons revealed that the average ratio of beer consumed to body weight in the positive condition was significantly greater than the average ratio in the neutral condition, and the average ratio of beer consumed to body weight was significantly less in the negative condition than the average ratio in the neutral condition. These findings are discussed as they relate to cognitive models of alcohol use.

Molecular evidence for distinct genotypes of monkey B virus (herpesvirus simiae) which are related to the macaque host species.

Although monkey B virus (herpesvirus simiae; BV) is common in all macaque species, fatal human infections appear to be associated with exposure to rhesus macaques (Macaca mulatta), suggesting that BV isolates from rhesus monkeys may be more lethal to nonmacaques than are BV strains indigenous to other macaque species. To determine if significant differences that would support this supposition exist among BV isolates, we compared multiple BV strains isolated from rhesus, cynomolgus, pigtail, and Japanese macaques. Antigenic analyses indicated that while the isolates were very closely related to one another, there are some antigenic determinants that are specific to BV isolates from different macaque species. Restriction enzyme digest patterns of viral DNA revealed marked similarities between rhesus and Japanese macaque isolates, while pigtail and cynomolgus macaque isolates had distinctive cleavage patterns. To further compare genetic diversity among BV isolates, DNA sequences from two regions of the viral genome containing genes that are conserved (UL27 and US6) and variable (US4 and US5) among primate alphaherpesviruses, as well as from two noncoding intergenic regions, were determined. From these sequence data and a phylogenetic analysis of them it was evident that while all isolates were closely related strains of BV, there were three distinct genotypes. The three BV genotypes were directly related to the macaque species of origin and were composed of (i) isolates from rhesus and Japanese macaques, (ii) cynomolgus monkey isolates, and (iii) isolates from pigtail macaques. This study demonstrates the existence of different BV genotypes which are related to the macaque host species and thus provides a molecular basis for the possible existence of BV isolates which vary in their levels of pathogenicity for nonmacaque species.

Shedding and transmission of baboon Herpesvirus papio 2 (HVP2) in a breeding colony.

Baboons in a captive breeding colony were monitored twice a year, and new additions were screened on arrival for shedding of Herpesvirus papio 2 (HVP2) and serologic reactivity to the agent. For 128 individual animals tested over a period of 1.5 years, shedding of infective virus was detected in 13 of 342 swab specimens (3.8%), each of these incidents representing shedding by a different animal. Among long-term colony animals, infective virus was recovered on only two occasions (5 of 236 swab specimens from five individuals). In all but one instance, animals shedding virus were infants, not adults, and all animals were shedding virus in the oral cavity. One of these five instances was an isolated case, but four (three infants and one adult) were clustered within a single breeding group. Molecular analyses of the HVP2 isolates from this cluster indicated that they likely arose from a single common source, probably the mother of one of the infants. None of 31 wild-caught baboons added to the colony during this period were found to be shedding infective virus, despite 93.5% of them being seropositive for HVP2. In contrast, 6 of 18 adult baboons (all seropositive) transferred into the colony from another breeding colony were found to be shedding HVP2 either orally (3 of 6) or genitally (3 of 6). In addition, 2 of 8 juvenile baboons in this shipment were found to be shedding virus in the oropharynx. Overall, 10 of 13 instances of HVP2 isolation were from the oropharynx rather than the genital tract, and 6 of 13 baboons shedding virus were infants or juveniles rather than adults. These results suggest that, although venereal transmission of HVP2 occurs among adult animals, oral infection of young, sexually immature baboons is not uncommon.

Detection and differentiation of primate alpha-herpesviruses by PCR.

A rapid method for detection and differentiation of 5 primate alpha-herpesviruses (human herpes simplex virus types 1 and 2 [HSV1, HSV2], green monkey simian agent 8, baboon herpesvirus 2 [HVP2], and macaque B virus [BV]) was developed utilizing the polymerase chain reaction (PCR). PCR primers were located in conserved regions of the gene encoding the glycoprotein B, which flanks an intervening region that is highly divergent among the 5 viruses. Amplified PCR products from the 5 viruses were readily differentiated by their unique restriction enzyme digestion patterns. No variation in digestion patterns was noted among strains of HSV1, HSV2, or HVP2. One clinical isolate of BV exhibited variation in a single restriction site, but its overall restriction pattern remained typical of BV. This method (PCR/RFLP) allowed the presence of herpesvirus DNA in clinical swabs from primates to be readily detected and the virus unambiguously identified.

Prevalence of Herpesvirus papio 2 in baboons and identification of immunogenic viral polypeptides.

The prevalence of Herpesvirus papio 2 (HVP2) in several groups of captive and wild-caught baboons was determined by detection of anti-HVP2 antibodies in 133 sera of adult baboons. Over 90% of newly imported (wild-caught) adult olive baboons (Papio anubis) from Kenya and chacma baboons (P. ursinus) from South Africa were found to have anti-HVP2 titers. Similarly, approximately 85% of captive breeding colony baboons (P. anubis and P. cynocephalus) were seropositive for HVP2. Infected animals were generally easily identifiable by enzyme-linked immunosorbent assay because anti-HVP2 IgG titers in immune animals were usually high (16,000 to 64,000). There was little variation in the relative reactivity patterns of individual HVP2-immune sera when tested against herpes simplex viruses 1 and 2, monkey B virus, H. cercopithecus 2, and HVP2, or against different HVP2 strains. Also, differences were not detected between reactivity of olive and chacma baboon immune sera. Analysis of the polypeptide specificity of immune sera by western blot identified four viral antigens that were consistent targets of immune sera. These antigens were the gB glycoprotein, a pair of unidentified glycoproteins of 80 to 100 kDa, the gD glycoprotein, and a series of smaller capsid proteins. Additional viral proteins were variably recognized by individual immune sera. The results of this study indicate that HVP2 is a common infection of baboons; there is little antigenic variation among HVP2 strains; and there are several HVP2 antigens that represent consistent targets of the anti-HVP2 immune response of baboons.

Herpesvirus papio 2, an SA8-like alpha-herpesvirus of baboons.

Several SA8 isolates obtained from baboons were compared to the prototype SA8 herpesvirus of African green monkeys. SDS-PAGE and restriction enzyme analyses revealed definite differences between green monkey and baboon isolates. DNA and amino acid sequences of the gB, gD and gJ glycoprotein genes exhibited substantial differences in variable regions. For the gB and gD, the amount of amino acid substitutions between SA8 and the baboon viruses was comparable to levels observed between analogous genes of SA8 & B virus or HSV1 & HSV2. Although a high degree of antigenic cross-reactivity was apparent, virus-specific antigenic determinants were also readily detected. Phylogenetic analyses supported separation of the baboon isolates and SA8 as distinct viruses. Taken together these results suggest that although closely related to SA8, the baboon viruses represent a distinct simian alpha-herpesvirus which we propose be designated Herpesvirus papio 2.

Gene mapping and sequence analysis of the unique short region of the simian herpesvirus SA 8 genome.

A 10.5 kbp BamHI restriction fragment representing most of the unique short (Us) region of the genome of the simian alpha-herpesvirus SA8 was identified and cloned. Partial sequencing of this DNA fragment identified regions of sequence homology with eight open reading frames (ORFs) of HSV1 and/or HSV2. Sequence and size analysis of subcloned fragments of the SA8 Us region and comparison with homologous HSV Us sequences determined that the number, order, size, and orientation of SA8 Us ORFs are comparable to those of HSV. Based on the location of transcriptional control elements, transcription of SA8 Us genes appears to be organized into 3' co-terminal mRNA sets as in HSV, although the grouping of the gene sets is different. The SA8 US4 (gG) ORF is more similar to that of HSV2 than HSV1, both in size and predicted amino acid sequence. Complete sequences were determined for five SA8 genes which represent homologs of the HSV gD, gE, gI, US5, and US9 genes. The predicted polypeptides encoded by SA8 are similar to the corresponding HSV polypeptides. All SA8 Us genes were more closely related to those of HSV than to related gene homologs of other mammalian alpha-herpesviruses.