Short communication: Dairy bedding type affects survival of Prototheca in vitro.

Protothecae are algal pathogens, capable of causing bovine mastitis, that are unresponsive to treatment; they are believed to have an environmental reservoir. The role of bedding management in control of protothecal mastitis has not been studied. The purpose of this study was to evaluate the growth of either environmental or mastitis-associated Prototheca genotypes in dairy bedding materials that are commonly used in Maine. Prototheca zopfii genotypes 1 and 2 (gt1 and gt2) were inoculated into sterile broth only (control ), kiln-dried spruce shavings, "green" hemlock sawdust, sand, or processed manure-pack beddings with broth, and incubated for 2 d. Fifty microliters of each isolate was then cultured onto plates and the resulting colonies counted at 24 and 48 h postinoculation. Shavings were associated with significantly less total Prototheca growth than other bedding types. Growth of P. zopfii gt1 was significantly higher than that of gt2 in the manure-pack bedding material. Spruce shavings, compared with manure, sand, or sawdust, may be a good bedding type to prevent growth of Prototheca. Based on these in vitro findings, bedding type may affect Prototheca infection of cattle in vivo.

Vaginal myeloperoxidase and flora in the pig-tailed macaque.

Myeloperoxidase (MPO) is an enzyme in neutrophils and monocytes which reacts with H2O2 and chloride to kill microbes after phagocytosis. Instillation of MPO into the vagina may augment vaginal defenses against sexually transmitted diseases, since the normal vaginal flora is characterized by the presence of H2O2-producing lactobacilli. We assessed the menstrual cycle stage, vaginal flora, pH, macroscopic appearance, and endogenous MPO in the adult female pig-tailed macaque (Macaca nemestrina) at baseline (n = 26; 60 observations) and at 0, 4, and 24 hours in untreated animals (n = 6) or in animals treated with intravaginal MPO gel at time 0 (n = 5). Baseline MPO levels were highly variable, and there was no detectable effect of cycle stage. In untreated animals, there was no significant effect of vaginal swab collection on vaginal flora or MPO levels. MPO treatment did not reduce vaginal H2O2-producing organisms, and vaginal MPO levels tended to increase at 4 hours in treated animals. Vaginal/cervical colposcopic changes were not detected in either group.

Antibody response to the chlamydial heat-shock protein 60 in an experimental model of chronic pelvic inflammatory disease in monkeys (Macaca nemestrina).

A primate model of chlamydial pelvic inflammatory disease was used to characterize serum antibody responses to the 60 kDa chlamydial heat shock protein (CHSP60). Forty monkeys were infected in the fallopian tubes with Chlamydia trachomatis and then were treated. Twenty-three (58%) monkeys developed antibodies against CHSP60, of whom 6 (15%) had CHSP60 responses that persisted throughout the study and 17 (42.5%) had a transient response. A persistent CHSP60 antibody response was correlated with being culture- or ligase chain reaction-positive in the fallopian tubes (P=.004), but not in the cervix pretreatment, and with being tubal-positive posttreatment (P=. 02). Compared with tubal-negative monkeys, tubal-positive monkeys had more intense CHSP60 responses (P=.006) that lasted longer (P=. 002). Among CHSP60 responders, an OD>0.5 was correlated with more severe salpingeal pathology before treatment (P=.04). CHSP60 antibody response may be useful as a marker of persistent chlamydial infection in the fallopian tubes.

Evidence of genetic susceptibility to Chlamydia trachomatis-induced pelvic inflammatory disease in the pig-tailed macaque.

The macaque model of chlamydial pelvic inflammatory disease (PID) demonstrates individual variability in the time of onset of intrapelvic adhesions. Some animals develop adhesions rapidly, within 2 weeks after a single tubal inoculation with Chlamydia trachomatis, while in others, adhesions are not observed until 2 weeks after a second tubal inoculation. To test whether this variability correlates with major histocompatibility complex (MHC) class I haplotype, we used macaque alloantisera and mouse anti-HLA monoclonal antibodies to determine the MHC class I haplotypes of 44 C. trachomatis-infected macaques (Macaca nemestrina). Macaques developing gross tubal adhesions after the first chlamydial inoculation were classified as susceptible (n = 29), while those not developing adhesions until after the second chlamydial inoculation were classified as relatively resistant (n = 15), to adhesion formation. Three antibody specificities correlated with susceptibility (odds ratio [OR] 5.2, P < 0.01; OR 6.1 and 4.3, P < 0.05), and two correlated with relative resistance to adhesions (OR 0.1, P < 0.05; OR 0.2, P < 0.01). Because several of these antibodies are cross-reactive, as many as five different MHC class I alleles (three increasing and two decreasing ORs) or as few as two different MHC class I alleles (one increasing and one decreasing OR) could be correlated with risk of adhesion formation. We conclude that in macaques, susceptibility or relative resistance to rapid formation of tubal adhesions is correlated with expression of MHC class I alleles, consistent with reports of MHC class I restriction of chlamydial immunopathology in humans.

Ejaculatory pattern of llamas during copulation.

The objective of this study was to use transrectal digital palpation of urethral pulses to define the ejaculatory pattern of llamas during copulation. Five male llamas were palpated during 5 to 6 copulations each with receptive female llamas (n = 28 copulations). The time from first exposure of a male to a female until mounting was 0.7 +/- 1.1 min (mean +/- SD), time to the first intromission was 1.7 +/- 1.4 min, and time from initial mount to final dismount (copulation duration) was 21.7 +/- 7.8 min. A total of 121.9 +/- 61.0 urethral pulses per copulation (5.6 +/- 1.7 pulses/min) was palpated. During the first 3.9 +/- 3.7 min of copulation, urethral pulses (11.0 +/- 10.1 urethral pulses at 3.5 +/- 2.5 pulses/min) occurred randomly and were not associated with whole-body strains. After the first 4 min of copulation, urethral pulses occurred in a pattern of clusters of frequent urethral pulses associated with whole-body strains, alternating with intercluster intervals of infrequent urethral pulses without whole-body strains. Individual clusters were characterized by 4.3 +/- 2.7 urethral pulses at 16.7 +/- 4.5 pulses/min during strains, and intercluster intervals were characterized by 1.7 +/- 2.3 urethral pulses at 2.2 +/- 1.8 pulses/min. Each cluster of urethral pulses during a strain was preceded by 2.3 +/- 1.8 repositions of the male's hindlegs and by 38.1 +/- 20.8 pelvic thrusts. There were 18.5 +/- 10.6 clusters of urethral pulses accompanied by strains per copulation at 0.9 +/- 0.3 clusters/min. The 18 to 19 clusters of urethral pulses appeared to be individual ejaculations. Therefore, we hypothesize that llamas ejaculated 18 to 19 times during their 22-min copulations.

Seminal collection, seminal characteristics and pattern of ejaculation in llamas.

Semen was collected from 10/10 llamas during 26/30 (87%) collection attempts using an artificial vagina mounted inside a surrogate female. For the 26 semen collections, the duration of copulation (mount to dismount) with the artificial vagina was 31.7 +/- 12.0 min (mean +/- SD). Seminal pH was 8.1 +/- 1.1, and seminal volume per collection was 3.0 +/- 1.9 ml. Sperm concentration per collection was 1.0 +/- 0.8 x 10(6) sperm/ml, total number of spermatozoa was 2.9 +/- 3.1 x 10(6), total sperm motility was 23.7 +/- 20.0%, and the percentage of morphologically normal spermatozoa was 39.7 +/- 18.5%. Morphologically abnormal spermatozoa were categorized according to abnormal heads (20.1 +/- 19.9%), tail-less heads (8.7 +/- 8.9%), abnormal acrosomes (12.9 +/- 12.4%), abnormal midpieces (1.0 +/- 3.7%), cytoplasmic droplets (11.1 +/- 12.4%), and abnormal tails (6.6 +/- 12.0%). There were 0.3 +/- 0.3 million motile, morphologically normal spermatozoa per collection: less than 1000 during the first 5 min of copulation, 0.01 +/- 0.01 x 10(6) between 5 and 10 min of copulation, 0.04 +/- 0.08 x 10(6) between 10 and 15 min of copulation, 0.09 +/- 0.21 x 10(6) between 15 and 20 min of copulation, and 0.15 +/- 0.28 x 10(6) between 20 min and the end of copulation.

Corpus luteal function in nonpregnant mares following intrauterine administration of prostaglandin E(2) or estradiol-17beta.

The objective of this study was to test the hypothesis that intrauterine administration of prostaglandin E(2) (PGE(2)) or estradiol-17beta (E-17beta) would prolong CL function in nonpregnant mares. Nonpregnant mares were continuously infused with 240 mug/d of PGE(2), 6 mug/d of E-17beta, or vehicle (sham-treated) on Days 10 to 16 post ovulation (ovulation = Day 0), using osmotic minipumps surgically placed into the uterine lumen on Day 10 (n = 11 per group). Nonpregnant and pregnant mares served as negative and positive controls, respectively (n = 11 per group). Mares were defined as having prolonged CL function if plasma progesterone remained > 2.5 ng/ml and if ovulation did not occur on Days 9 to 30. Corpus luteal function was prolonged until Day 30 in 1 11 nonpregnant mares, 4 11 sham-treated mares, 6 11 E-17beta-treated mares, 8 11 PGE(2)-treated mares, and 11 11 pregnant mares. The incidence of prolonged CL function was similar (P=0.16) in the sham-treated and nonpregnant mares. The hypothesis that PGE(2) would prolong CL function in nonpregnant mares was supported, since the incidence of prolonged CL function was higher (P=0.003) in PGE(2)-treated versus nonpregnant mares, tended to be higher (P=0.09) in PGE(2)-versus sham-treated mares, and was not lower (P=0.11) in PGE(2)-treated versus pregnant mares. The hypothesis that E-17beta would prolong CL function in nonpregnant mares was not supported, since the incidence of prolonged CL function was not higher (P=0.34) in E-17beta-versus sham-treated mares, and was lower (P=0.02) in E-17beta-treated versus pregnant mares. These results demonstrate that intrauterine administration of a pharmacologic dose of PGE(2) initiated prolonged CL function in nonpregnant mares. Further experiments are needed to confirm the role of conceptus secretion of PGE(2) in CL maintenance, and to determine the mechanism of action of PGE(2) within the equine reproductive tract.

Uterine transport of prostaglandin E(2)-releasing simulated embryonic vesicles in mares.

Transrectal ultrasonography was used to test the hypothesis that prostaglandin E(2) (PGE(2)) would increase the uterine transport of simulated embryonic vesicles in mares. Uterine transport of PGE(2)-releasing (PGE) vesicles, vehicle-releasing (sham) vesicles, and equine embryos was contrasted on Day 12 or Day 13 post ovulation. In Experiment 1, there was no difference (P>0.10) in transport of PGE vesicles, sham vesicles, Day-12 embryos, and Day-12 embryos after cervical manipulation (n = 3 per group). In Experiments 2 and 3, respectively, transport of PGE and sham vesicles was contrasted with transport of Day-13 embryos after the vesicles (1 vesicle per mare) were placed into the uterine lumen with the embryo, (n = 7 per group). In Experiment 2, PGE vesicles were transported less often (P<0.05) from horn to body and from segment to segment than Day-13 embryos before vesicle insertion. In Experiment 3, sham vesicles were transported less often from horn to body (P<0.10) and from segment to segment (P<0.01) than Day-13 embryos before vesicle insertion. However, there was no difference (P>0.10) in the transport of PGE vesicles and embryos (Experiment 2) or sham vesicles and embryos (Experiment 3) together in the uterine lumen. In Experiment 4, transport of PGE and sham vesicles was contrasted by placing them together into the uterine lumen of nonpregnant mares on Day 13 (n = 7). There was no difference (P>0.10) in the transport of PGE and sham vesicles together in the uterine lumen. These results do not support the hypothesis that PGE(2) increases uterine transport of simulated embryonic vesicles. In addition, these results do not support the hypothesis that equine embryos stimulate uterine transport.

Distribution of bovine cysticercosis in Washington.

Data from slaughter plants (n = 3) and feedlots (n = 18) in eastern Washington were analyzed to characterize occurrence patterns of cysticercosis in Washington during 1984. Three concurrent peaks in cysticercosis rates (0.6/1,000 to 5/1,000 slaughtered cattle) were detected at 3 slaughter plants. Peaks were observed at 8 feedlots from December 1983 to March 1984, at 6 feedlots from April to July 1984, at 2 feedlots from August to October 1984, and at 3 feedlots from November 1984 to February 1985. Affected feedlots were not closely associated geographically and were feeding cattle from many, predominantly northwestern, origins. For 3 feedlots for which time in the feedlot was available for each slaughter shipment, an increase in cysticercosis rate with increasing time in the feedlot was noticed. Within these 3 feedlots, cases of cysticercosis were widely scattered spatially. The pattern of cysticercosis indicated human fecal contamination of a regionally available feed source. Of feedstuffs in use, potato waste, a byproduct of the processed potato industry, appeared to be the most likely source of Taenia saginata ova.