Bacteriological, cytological and histopathological evaluation of the reproductive tract of slaughtered cows / Avaliação bacteriológica, citológica e histopatológica do trato reprodutivo de vacas abatidas

Reproductive diseases, mainly endometritis, are important hurdles in cattle raising, In the current study we evaluated gross, bacteriological, cytological, and histological findings from selected sites of the genital from 23 slaughtered cows and tested whether there is an association between these findings and the probability of reaching a reliable diagnosis. The results from the examinations of macroscopic aspects of uterine secretions, the cytological, bacteriological, and histopathological findings were then correlated. There was no significant correlation (P>0.05) of the statistical data from different parts of the genital tract. Trueperella pyogenes and Escherichia coli were isolated from the vagina in 3/23 cases. In only 2/23 samples Enterococcus faecalis and a gram-negative, oxidase-positive bacteria were isolated from the cervix uteri. Only Staphylococcus epidermidis, in 1/23 case, was isolated from the uterus. Histopathological findings in uterus from samples of Groups II (moderate lesions) and III (severe lesions) did not translated in grossly visible changes. Samples from reproductive tracts with secretion in the vagina and cervix uteri had no detectable changes in the other parameters analyzed from this portion. Uterus with positive bacterial culture had evidence of ascendant inflammation judging by the high granulocyte count in the three analyzed portions. This study reinforces that vaginitis and cervicitis in the cow diagnosed only by clinical examination does not reflect the real status of the uterine health. For this reason, treatment of uterine disorders should be conducted based on reliable tests to determine the appropriate therapy for each situation.(AU)

Doenças reprodutivas causam altas perdas econômicas nos rebanhos bovinos, sendo a endometrite uma das alterações mais relevantes. Os objetivos desta pesquisa foram avaliar a associação dos achados fisiopatológicos em exames macroscópicos, bacteriológicos, citológicos e histopatológicos nas porções do trato genital de 23 vacas abatidas, bem como avaliar a necessidade de associação destes exames para efetuar diagnóstico fidedigno. A avaliação macroscópica da secreção, os exames histopatológico, citológico e as bactérias identificadas foram correlacionados. Não houve associação (P>0,05) dos resultados nas diferentes porções do trato genital. Na vagina foram isoladas as bactérias Trueperella pyogenes e Escherichia coli. Na cérvix, em apenas 2/23 (8,6%) amostras isolou-se Enterococcus faecalis e gram negativo oxidase positiva. No útero houve isolamento apenas da bactéria Staphylococcus epidermidis. As amostras histopatológicas classificadas em grupo II e III não apresentaram alterações detectadas no exame macroscópico. As amostras com secreção não fisiológica na vagina e cérvix não apresentaram alterações nos outros exames na porção uterina. A amostra com cultura bacteriológica positiva no útero demonstrou uma infecção ascendente através da alta contagem de granulócitos nas três porções analisadas. O presente estudo reforça o conceito de que a vaginite e cervicite diagnosticadas pelo exame clinico na vaca não retratam o real status da saúde uterina e por esta razão, o tratamento do útero deve ser conduzido com critério, alicerçado nos exames complementares para definir a terapia adequada para cada situação.(AU)

Prevalence of cervicitis in dairy cows and its effect on reproduction.

The objective of this study was to determine whether cervicitis in dairy cows is an independent disease or occurs concomitantly with inflammation of the uterus, and to clarify possible effects of cervicitis on reproductive performance. Dairy cows (n = 416) from 33 dairy farms were examined by rectal palpation and vaginoscopy between 42 and 50 days postpartum. Inclusion criteria for this study were absence of abnormal vaginal discharge and abnormalities of the uterus (fluctuation) at rectal palpation. Cervicitis was diagnosed when the second cervical fold was swollen and prolapsed with (C2) or without (C1) reddening. Cytobrush samples from the uterus (n = 370) and the cervix (n = 402) were collected, and the percentage of neutrophils in the uterus (PMNU) and the cervix as indicators of inflammation (threshold: ≥5%) was determined. In addition, endometrial biopsies for histology were collected, 300 of which were suitable for evaluation. Cervicitis (C1/C2) was diagnosed in 253 of 416 (60.8%) of cows. Of these, the prolapsed cervical mucosa was hyperemic (C2) in 29.1% of cases. Of 370 available uterine cytology samples, 221 cows had a clinical cervicitis; however, 170 (76.9%) had PMNU less than 5%. Of 300 uterine histologic examinations, 82 (27.3%) did not reveal any abnormalities; the remaining cows either had uterine inflammation and/or degenerative uterine changes such as endometriosis and angiosclerosis. Furthermore, of 300 biopsied animals, 184 revealed a cervicitis (C1/C2); however, 30.4% of these animals had no histopathologic uterine findings. For further analysis, only animals either without histopathologic findings and normal uterine cytology or with solely endometritis (defined as PMNU ≥ 5% and/or positive histopathology of the uterine tissue) were evaluated (n = 157). Of these, 95 cows had cervicitis. Unexpectedly, 63 of 95 (66.3%) cows had cervicitis without endometritis. With regard to reproductive performance, days to first service were not affected by cervicitis. Number of days open in animals with cervicitis but without endometritis tended to be lower than in cows with cervicitis plus endometritis (P = 0.092). Also, number of days open relative to percentage of neutrophils greater than 5% was lower when the cervical compared to the uterine mucosa was affected (P < 0.05). Total conception and pregnancy rates of animals 200 days into lactation decreased significantly in cows with severe cervical inflammation (C2). In conclusion, the results of this study suggested that cervicitis occurs independent of endometritis, and a higher degree of cervicitis is associated with poorer reproductive performance.

Interactions of metabolism, inflammation, and reproductive tract health in the postpartum period in dairy cattle.

This paper reviews recent data and concepts on the development of inflammation in the reproductive tract of dairy cows during the first 2 months after calving. The incidence of metritis is typically 10-20%, with 5-15% of cows having purulent vaginal discharge (PVD), 15-40% having cervicitis approximately 1 month after calving, and 10-30% having cytological endometritis between 1 and 2 months after calving. Endometritis, cervicitis and PVD are distinct conditions, each of which is associated with significantly increased time to pregnancy, and affected cows often have more than one of these conditions. Cumulatively, 35-50% of cows have at least one form of pathological reproductive tract inflammation between 3 and 7 weeks postpartum. It is hypothesized that reproductive tract disease represents a failure of the immune system to switch fast enough or far enough from the down-regulated state necessary for maintenance of pregnancy to a heightened state of function for postpartum clearance of bacteria and tissue debris and then to a 'quiet' state 3-4 weeks later. There are numerous links between fat metabolism, inflammation and immune function, and changes in these precede reproductive tract disease by several weeks. An excessive pro-inflammatory state early in the postpartum period appears to be a key feature of cows with endometritis approximately 1 month later. Generally, worse postpartum negative energy balance (NEB) is associated with more severe or prolonged uterine inflammation. Aspects of both mononuclear cell proliferation and neutrophil oxidative burst are commonly impaired, particularly in association with elevated non-esterified fatty acid concentrations and to a lesser degree by ketosis. In summary, NEB contributes to immune dysfunction which in turn is a major component of reproductive tract inflammatory disease. The factors that initiate and sustain harmful inflammation of the reproductive tract are not yet well quantified.

Effect of endocervical inflammation on days to conception in dairy cows.

In contrast to endometritis, now diagnosed by cytological examination, the effect of endocervical inflammation on reproductive performance has been inadequately investigated. In this study, endocervical and endometrial cytological specimens were collected from 168 Holstein cows between 21 and 60 days in milk (DIM) to investigate the prevalence of endocervical inflammation and effect on days to conception. Statistical analyses were stratified based on DIM at examination (<35 vs. ≥35 DIM). Endocervical inflammation with ≥5% neutrophils before 35 DIM (disregarding the level of endometrial inflammation) was associated with decreased hazard of pregnancy within 300 DIM (adjusted hazard ratio, 0.4; 95% confidence interval: 0.3-0.8). A decrease in hazard of pregnancy was observed when >6% neutrophils were counted in endometrial smears (adjusted hazard ratio, 0.4; 95% confidence interval: 0.2-0.7). The study suggested an additive effect of combined endocervical and endometrial inflammation on the hazard of pregnancy within 300 DIM. Using the thresholds of 5% neutrophils for the cervix and 6% neutrophils for the uterus, 11% of the cows examined before 35 DIM presented cervicitis only, 13% were affected by endometritis only, and 32% suffered from both endometrial and endocervical inflammation. The presence (absence) of cervicitis was not indicative of the presence (absence) of endometritis. This study showed that in addition to uterine inflammation, endocervical inflammation in early lactation affects conception. Thus, the global evaluation of genital tract health may be more beneficial for reproductive performance than that of endometrial inflammation.

In captive rhesus macaques, cervicovaginal inflammation is common but not associated with the stable polymicrobial microbiome.

Vaginal inoculation of rhesus macaques (RM) with simian immunodeficiency virus (SIV) has been used to study the biology of HIV transmission. Although the results of vaginal SIV transmission experiments could be affected by vaginal inflammation, studies to date have been conducted without regard to levels of pre-existing genital inflammation present in RM. We collected cevicovaginal secretions (CVS) from 33-36 RM during the mid menstrual cycle (day 10-20) at 2 time points approximately 8 months apart and characterized the mRNA and protein levels of inflammatory cytokines, chemokines and interferon-stimulated genes. There was extreme variability in the levels of inflammatory mediators (IFN-α, IFN-γ, IL-6, TNF, IL-1b, IP-10, MIG, IL-12 and IL-17). In most animals, the mRNA levels of the inflammatory mediators were similar in the 2 CVS samples collected 8 months apart, suggesting that genital inflammation is stable in a subset of captive female RM. At both time points the cervicovaginal microbiota had low levels of Lactobacillus and was relatively diverse with an average of 13 genera in the samples from the first time point (median 13, range 7-21) and an average of 11.5 genera in the samples from the second time point (median 11, range 5-20). Many of the macaques had similar microbiota in the samples collected 8 months apart. However, we found no correlation between specific bacterial genera and the mRNA or protein levels of the inflammatory mediators in the genital tract of RM in this study. It seems likely that results of published vaginal SIV transmission experiments in RM have been influenced by pre-existing inflammation in the animals used for the experiments.

Isolation of Haemophilus somnus from dairy cattle in kwaZulu-Natal. An emerging cause of 'dirty cow syndrome' and infertility?

Haemophilus somnus was consistently isolated from vaginal discharges of dairy cows submitted from field cases of vaginitis, cervicitis and/or metritis in the KwaZulu-Natal Midlands during the period July 1995 - December 2000 and from the East Griqualand area in November/December 2000. The purulent vaginal discharges, red granular vaginitis and cervicitis, and pain on palpation described in these cases was very similar to that reported in outbreaks of H.somnus endometritis syndrome in Australia, Europe and North America. In all the herds involved in these outbreaks, natural breeding with bulls was employed. Although there was a good cure rate in clinically-affected animals treated with tetracyclines, culling rates for chronic infertility were unacceptably high. Employment of artificial insemination in these herds improved pregnancy rates in cows that had calved previously, but many cows that had formerly been infected failed to conceive.

Hysterotomy by a colpotomy approach for treatment of foetal mummification in a cow.

A 7-year-old Brahman cow was diagnosed as suffering from chronic foetal mummification of unknown aetiology, concurrent cystic ovarian disease, prolapse of the 2nd cervical ring and chronic cervicitis. Repeated treatment with prostaglandin F2alpha and oestrogen failed to resolve the mummification. A hysterotomy was performed via an incision in the dorsolateral vaginal wall. Good exposure of the uterine horn was achieved and mild post-operative complications were observed. Colpotomy can be regarded as an alternative surgical approach to the moderately enlarged bovine uterus.

Alteraçöes morfológicas em ovários e úteros de ovelhas (Ovis aries) / Morphologic alterations in the ovaries and uterus of sheep (Ovis aries)

Relatam-se os achados anátomo-histopatológicos em ovários e úteros de 225 ovelhas lanadas, adultas e näo-gestantes colhidos no Estado do Rio Grande do Sul - Brasil, sendo 117 animais procedentes do Município de Alegrete (Grupo I) e 108 do Município de Bagé (Grupo II). Os resultados indicaram a existência de 113 (96,58 por cento) animais com alteraçöes morfológicas no Grupo I e 96 (88,89 por cento) no Grupo II. A distribuiçäo de acordo com a natureza das alteraçöes, respectivamente para os Grupos I e II, foi: do desenvolvimento - 2,56 por cento e ausentes; regressivas - 52,99 por cento e 22,22 por cento; inflamatórias - 55,56 por cento e 79,63 por cento e progressivas - 146,15 por cento e 91,67 por cento. O comprometimento da reproduçäo, embora näo corresponda quantitativamente ao elevado volume de patologias, alcança expressividade nos animais estudados. Os transtornos da reproduçäo, representados principalmente pelos processos inflamatórios uterinos atingem os valores respectivos para os Grupos I e II de 63,24 mais ou menos 8,75 por cento (infertilidade - 11,97 mais ou menos 5,6 por cento; subfertilidade - 51,28 mais ou menos 9,01 por cento) e 66,67 mais ou menos 8,9 por cento (infertilidade - 9,26 mais ou menos 5,4 por cento; subfertilidade - 57,41 mais ou menos 9,37 por cento). Verificou-se, que os animais mais jovens apresentaram um maior percentual de alteraçöes inflamatórias. A tendência à simultaneidade da endometrite e cervicite foi significativa (P<0,05)

[Detection of Chlamydia in inflammation of the cervix and uterus in cows]. / Nálezy chlamydií u zánetu delozního krcku a pochvy krav.

Clinical findings in cows of seven farms with a frequent occurrence of birth canal inflammations were matched against results of serological and direct immunofluorescence tests for Chlamydia sp. infection. Normal clinical picture, weak to moderate and serious inflammations were found in 16.1%, 46.7% and 37.2% of the 205 examined cows, respectively. The results of micro CFT were negative in 32.7% and titres 1:5, 1:10, 1:20, and 1: > or = 40 were found in 8.7%, 38.0%, 12.3%, and 8.3%, respectively. Direct immunofluorescence test of cervical swabs, using the Progen Biotechnic GmbH set, was negative in 29.1% and clearly positive in 34.7% of the tested cows. The rest was animals in which sporadic elementary bodies were seen. The significance of the findings is discussed and the necessity to test Chlamydia infections in herds affected by frequent and/or chronic inflammations of genital organs is stressed. Evidence of the transmission of Chlamydia infection by contaminated semen is presented.

Therapeutic efficacy of povidone-iodine (Betadine) and dichloroxylenol (Septocid) in Holstein cows affected with endometritis and/or cervicitis.

The efficacy of either povidone-iodine (Betadine) or dichloroxylenol (Septocid) intrauterine infusions on the treatment of endometritis and/or cervicitis in cows was examined. One hundred and twelve repeat-breeder Holstein cows (aged 3-7 years) were selected for this study. Rectal and vaginal examinations were applied to detect the signs of endometritis and/or cervicitis. Cows were assigned into two groups: the first group (n = 60) was treated with Betadine solution (0.5%) while the second group (n = 52) was given Septocid (0.1%) administered as intrauterine infusion (100-150 ml). Both treatment were repeated at least two times at 7-day intervals and the cows were rechecked. The responding animals were inseminated 10-12 h after oestrus detection using frozen semen from bulls of proven fertility. The success or failure of treatment was evaluated on the basis of the post-treatment conception rate. The relationship of the body weight of cows, the length of the service period (open days) and the number of previous services with the results of treatment with Betadine or Septocid is discussed. The recovery and conception rates obtained after Betadine treatment were better than those obtained after Septocid. Moreover, healthy cows (> or = 500 kg body weight) and those inseminated before post-partum day 180, having no more than 4-7 previous services, responded well to either Betadine or Septocid treatment.

Alteraçöes morfológicas em ovários e útero de vacas azebuadas näo gestantes. I. Alteraçöes inflamatórias: metrite, endometrite e cervicite / Ovarian and uterine pathological changes in cow. I. Metritis, endometritis and cervicitis

Estudou-se macroscópica e microscopicamente o útero de 168 vacas azebuadas, näo gestantes, procedentes de vários municípios do Estado do Mato Grosso do Sul. Do total de animais estudados, 96 (57,1 por cento) apresentaram alteraçöes inflamatórias no útero, assim distribuídas: seis (3,6 por cento) referentes a metrite, 11 (6,5 por cento) a cervicite e 80 (47,6 por cento) a endometrite

Morphometric studies of the bovine uterus: microscopic lesions and retrospective reproductive history.

A survey of reproductive tracts from nonpregnant cows at an abattoir was conducted. Reproductive tracts from 98 cows were studied microscopically and evaluated with a grading system to determine the severity of pathologic changes. Inflammatory reaction (type and distribution), fibrosis, gland morphology, and appearance were measured, quantified, and scored (1 to 4). Category 1 endometrium (normal) was found in 18 cows, category 2 in 23, category 3 in 34, and category 4 in 23. The category of endometrium was then compared with the retrospective reproductive status, and it was found that reproductive problems had occurred in 6 cows (33.3%) in category 1, in 13 cows (56.5%) in category 2, in 25 cows (73.5%) in category 3, and in 21 cows (91.3%) in category 4. Cervicitis was found in 43 (43.8%) genital tracts; 16 cows (16.3%) had salpingitis. Ovarian lesions were not observed. Serum samples were tested for certain antibodies. Only 1 was seropositive for brucellosis, but 61 of 81 sera had leptospira titers greater than 1:100 and 1:50 (hardjo). The serovar hardjo was the most prevalent. All sera had neutralizing titers against parainfluenza type 3 virus; 74.2% and 56.7% of the sera had neutralizing antibodies against bovine viral diarrhea virus and infectious bovine rhinotracheitis virus, respectively.

Infertility due to foreign objects in the cervix of a cow.

Two fetal claws were removed from the cervix of a Holstein cow that had a chronic vulvar discharge. The cow had calved 1 year earlier. After removal of the claws, localized peritonitis developed and the cervix remained open and draining.

Lesions of contagious equine metritis in mares.

Twenty-three mares were infected with contagious equine metritis organism by intrauterine inoculation, and necropsied after intervals of two to 116 days. Severe diffuse subacute salpingitis was seen in one mare, and mild multifocal subacute salpingitis was common. Severe diffuse endometritis and cervicitis initially were acute and became more severe, subacute and predominantly plasmacytic by 14 days, then declined but persisted as mild diffuse or multifocal inflammation for the rest of the experimental period. Vaginitis arose in parallel but resolved after 70 days. There were no lesions in the clitoral fossa or clitoral sinus.

Contagious equine metritis: effect of intrauterine inoculation of contagious equine metritis agent in pony mares.

Actively growing culture of contagious equine metritis (CEM) bacteria or infective exudate (or both) were inoculated intrauterine in pony mares. A direct relationship was observed between (i) appearance and duration of cervicitis and vaginitis and (ii) vaginal exudate. Clinical signs appeared 1 to 3 days after mares were inoculated and lasted 7 to 23 days. In the acute phase of infection, all uterine and cervical samples yielded CEM bacteria. In the asymptomatic stage of infection, CEM bacteria were not isolated from uterine and cervical samples; however, in 33%, 28%, and 20% of the pony mares, CEM bacteria were present in clitoral fossa, clitoral sinus, and urethral vestibule, respectively, Sampling during early estrus increased the bacterial isolation rate to 57% in mares that were previously negative; however, 3 days later, CEM bacteria could not be isolated from 62% of the positive mares. The results of repeated exposure experiments indicated the presence of local antibodies, as no CEM bacteria could be recovered at 2, 7, and 15 days after reexposure with a small number of bacterial cells (8.4 x 10(5) cells). The CEM bacteria were isolated from all mares reexposed with a large number of bacterial cells (7.2 x 10(8)) at 2 days after second inoculation and from 50% at 7 days. However, all of the mares were negative by day 15 after reexposure, indicating increased resistance to CEM bacteria.

Responses of pony mares to the agent of contagious equine metritis 1977.

Reproduction of contagious equine metritis 1977 in Pony mares was achieved with cultures of an unclassified Gram-negative coccobacillus. Infected mares developed a vaginal discharge and associated inflammatory changes of the cervix and vagina. There was evidence of variation in pathogenicity between different strains of the organism. Although all infected mares made spontaneous clinical recoveries, the Gram-negative coccobacillus persisted in the genital tracts of a considerable proportion for a variable period after challenge. Recovery of the organism was not associated solely with the occurrence of oestrus. None of the mares has carried over infection into the following breeding season. There was no evidence of localization of the organism in the urinary tract. Cytological examination of smears of cervical and urethral swabs was of diagnostic value only during the clinical phase of the infection. A serological response was demonstrable in all mares that became infected after exposure to the Gram-negative coccobacillus. The complement-fixation test gave more specific and clear-cut results than either the agglutination or the antiglobulin test, with which there was a problem with non-specific reactions. The experimental findings indicate the value of the complement-fixation test for confirming recent cases of contagious equine metritis in the mare.

Contagious equine metritis: clinical description of experimentally induced infection.

Contagious equine metritis was reproduced experimentally in 6 pony mares. The disease was characterized by rapidly developing cervicitis and by varying amounts of exudate. The exudate, apparent as early as 48 hours after inoculation, drained from the cervix as a tenacious, mucopurulent discharge for several days, then rapidly disappeared. In all mares, the clinical disease cleared within several weeks of inoculation, without treatment.