History Of Reproductive Disorders Research Articles

Seroepidemiological study of leptospirosis in horses with reproductive disorders

Leptospirosis is an anthropozoonosis caused by different species of bacteria of the genus Leptospira spp., resulting in economic losses in equine husbandry, including abortion, birth of weak and premature animals. In this context, the present study aimed to conduct a seroepidemiological study by identifying antibodies to Leptospira spp. in horses with a history of reproductive disorders. For this purpose, 108 horses from two studs in the Triângulo Mineiro region, designated A and B (A: n=68 and B: n=40), were subjected to the Microscopic Agglutination Test (MAT). Questionnaires were administered to the stud owners to analyze risk factors. Of the 108 samples analyzed, 50.9% (55/108) were reactive to MAT, with the most prevalent serovars being Djasiman 65.5% (36/55) and Hebdomadis 34.5% (19/55) with titers of 800 and 1600, respectively. Positivity was significantly higher (p<0.05) in females over 48 months of age in both studs A and B. In this context, it can be concluded that veterinarians should request leptospirosis laboratory tests in horses that present reproductive disorders, and sanitary measures should be recommended aiming to minimize economic losses as well as to avoid the potential zoonotic risk of the disease.

Characteristics of expression of proteomic markers in women with “thin” endometrium and their importance for the therapeutic effects of the use of exogenous estradiol: A prospective cohort comparative study

Aim. To analyze the relationship between the expression of proteomic markers [estrogen (ER) and progesterone (PR) receptors, LIF, FOXA1, FOXA2] of endometrial receptivity and indicators of the levels of sex steroids [estradiol (E2) and progesterone (P)] in the peripheral blood during the “implantation window” in women with a history of reproductive dysfunction with “thin” endometrium (TE) and evaluate their significance for the therapeutic effect of using estradiol drugs. Materials and methods. The prospective cohort comparative study was conducted. The main group included 52 patients with TE (7 mm according to ultrasound on preovulatory days), the comparison group consisted of 62 women with normal endometrial thickness (≥7 mm according to ultrasound), women of both groups with reproductive dysfunctions of unknown reason; the control group included 16 healthy women. Aspiration biopsy of the endometrium was performed on the 6–8 days after ovulation (LH +6–8), as well as venipuncture to obtain a sample of peripheral blood to determine the E2, P levels. A combined histological and immunohistochemical study of endometrial samples was performed (ER, PR, LIF, FOXA1, FOXA2). The therapeutic effects of estradiol drugs were assessed in a cohort of women from the main group with TE (n=16 out of 26, those who have not changed their reproductive plans). Results. All women had ovulatory values of progesterone [P≥16.1 nmol/l (LH +6–8)] and normoestrogenemia in the blood (p0.05). According to the results of the correlation analysis, there were no associations between the E2, P levels and the value of M-echo, and no relationships were found between the E2, P levels and the expression of ER, PR, LIF, FOX-proteins. It was revealed that there are significant positive relationships between M-echo and FOXA2 expression in the endometrium (r=0.422; p0.001). Conditional threshold values for the ER, PR expression (LH +6–8) were determined, the excess of which is associated with abnormal expression of LIF, FOX-proteins: for PR in the glands – 105 and 285, for ER in glands – 145, for ER in the stroma – 155. It was found that usual doses of estradiol drugs (1–1.5 mg/day) are capable of increasing endometrial proliferation in women with TE, provided that the expression of ER, PR, LIF, FOX-proteins in their endometrium is similar to healthy women. Increased doses of estradiol drugs (4 mg/day) in women with TE had a significant effect in the form of an increase M-echo value and/or pregnancy in cases where the endometrial expression of LIF, FOX-proteins was comparable with healthy women, even in the presence of hormone-receptor interactions in the endometrium that are different from healthy women. Spontaneous pregnancy in participants of the main group with hypoplastic endometrium in our study occurred in 5 women in whom the expression characteristics of FOXA1 and/or FOXA2 in the endometrium were comparable to healthy women. Conclusion. TE is not the only marker of an imbalance of proteomic molecules, but it determines a greater likelihood of impaired expression of ER, PR, LIF, FOX-proteins in the endometrium. In women with an unclear reason of fertility disorders, it is necessary to conduct an in-depth examination of endometrial samples. Characteristics of endometrial expression ER, PR, LIF, FOX-proteins and the usefulness of hormone-receptor interactions in the TE in women with a history of reproductive disorders can be markers for predicting the effectiveness of the use of exogenous estradiol drugs in women in order to enhance endometrial proliferation with an initially low M-echo.

Occurrence of Coxiellosis in ruminants and its associated risk factors

Coxiellosis in animals is caused by the zoonotic pathogen, Coxiella burnetii. Although the disease is of public health importance it remains underdiagnosed and underreported. The cross- sectional study was aimed to estimate the occurrence of the disease in livestock of study area and also to identify the risk factors associated with the disease in animals. Blood, serum, and vaginal swabs samples were collected from 200 ruminants (cattle, sheep, and goats), across various farms in Karnataka, India. These samples were then screened using ELISA and PCR (com1 and IS1111). A questionnaire was administered to the farm owners to collect the risk factor-related information. About 5.26 % cattle, 12.3 % sheep, and 12.5 % goats were positive by ELISA. By PCR, 9.47 % cattle, 9.3 % sheep, and 10 % goats were positive. Overall, the occurrence of 14.73 %, 18.46 % and 17.5 % was estimated in cattle, sheep and goat, respectively. PCR targeting the IS1111 gene detected higher number of samples as positive as compared to the com1 gene PCR. Higher number of vaginal swab samples were detected as positive as compared to blood. History of reproductive disorders (OR: 4.30; 95 %CI:1.95- 9.46), abortion (OR: 30.94; 95 %CI:6.30- 151.84) and repeat breeding (OR:11.36; 95 %CI:4.16- 30.99) were significantly associated with coxiellosis (p < 0.005). Multivariable analysis by logistic regression model analysis suggested retained abortion, repeat breeding and rearing of animal in semi-intensive system as factors significantly associated with the infection. Cultural identification of the PCR positive samples were cultured using embryonated egg propagation and cell culture techniques and positivity was confirmed in six samples. Phylogenetic analysis of the com1 and IS1111 gene revealed clustering based on similar geographic locations. The study estimated the occurrence of the disease in the study area and identified the potential risk factors.

Risk Factors Associated with Prevalence of Zoonotic Tuberculosis in Small Ruminants in Chhattisgarh, India

This study carried out in 2021-22 and 2022-23 at the College of Veterinary Science &amp; A.H., Anjora, Durg, and ICAR-NIBSM, Baronda, Raipur, Chhattisgarh, India focused on examining molecular detection and determining risk factors associated with infections of Bovine tuberculosis (BTB) in small ruminants within the Chhattisgarh region. Zoonotic tuberculosis, a global threat emerging from the Mycobacterium tuberculosis complex, impacts humans, livestock, and wildlife. A total of 795 samples from small ruminants, including blood, aborted fetuses, vaginal swabs, and placental tissues, were collected. DNA extraction and multiplex PCR targeting Mycobacteria RD1 and RD4 genes were performed. Statistical analyses explored individual animal-level and herd-level risk factors using SPSS. Results revealed a 0.75% prevalence of BTB in Small ruminants. Herd-level analysis demonstrated 2.6% prevalence. The statistical analysis assessing individual animal-level risk factors associated with BTB prevalence showed no significant associations with species, age, sex, or sample type. Herd-level risk factors included the number of small ruminants (OR=1.158, p =0.003) and introducing new animals (OR=1.090, p=0.034). In a multivariable logistic regression analysis, the variable “Number of small ruminants kept” emerged as a significant predictor (OR=1.155, p=0.004).Other factors like vaccination, biosecurity measures, and history of reproductive disorders did not reach statistical significance. This study provides comprehensive insights into the multifaceted risk factors associated with zoonotic tuberculosis transmission in small ruminants. The findings underscore the importance of ongoing research to formulate effective control strategies considering the nuanced nature of BTB.

Endometrial expression of FOX proteins (FOXA1 and FOXA2) in women of reproductive age with different endometrial thickness: prospective cohort comparative study

Aim. To evaluate the expression of FOX proteins (FOXA1 and FOXA2) in the endometrium during the implantation window in women with a history of reproductive dysfunctions with different thickness of the endometrium.&#x0D; Materials and methods. The prospective cohort comparative study was conducted. The main group included patients with "thin" endometrium (7 mm according to ultrasound on preovulatory days; n=52), the comparison group consisted of women with normal endometrial thickness (7 mm according to ultrasound; n=62; women of both groups with reproductive dysfunctions of unknown reason), the control group included 16 healthy fertile women. Aspiration biopsy of the endometrium was performed on the 68 days after ovulation, as well as venipuncture to obtain a sample of peripheral blood to determine the levels of sex steroids (estradiol E2 and progesterone P). A combined histological and immunohistochemical study of endometrial biopsies was performed.&#x0D; Results. All women had ovulatory values of progesterone P16.1 nmol/l (68 days after ovulation) and normoestrogenemia (E2, pmol/l) in the blood. E2/P was similar in all groups (p0, 05). A pronounced expression of FOXA1 was noted in women with thin endometrium significantly more often (p0.05) with various hormone-receptor characteristics of the endometrium (42% n=22 out of 52) compared with healthy participants (0%; n=0). Reduced FOXA2 expression in the uterine mucosa was significantly more often detected on 68 days after ovulation in women with "thin" endometrium (56% n=29 of 52) than in women with normal endometrial thickness, both in women from the comparison group and in healthy women from the control group (p0.05). In a generalized analysis of the expression of FOX proteins in the endometrium on days 68 after ovulation, it was generally found that every second (50%; n=57 out of 114) women with a history of reproductive disorders (with a reduced and normal M-echo value) expression of proteomic markers differed from healthy women. In the case of "thin" endometrium, more than two thirds of patients (71%; n=37 out of 52) showed differences in endometrial expression of FOX proteins compared with women without a burdened reproductive history.&#x0D; Conclusion. In the majority of women (71%) with a thin endometrium and a history of reproductive dysfunctions, the expression of FOX proteins in the endometrium differed from the control group. Overall, endometrial expression of FOX proteins, which is likely to be different from healthy women, in patients with reproductive dysfunctions of unknown origin is a significant predictor of reproductive failure. At the same time, such an isolated indicator as M-echo value 7 mm according to ultrasound data is not an absolute prognostic marker of endometrial receptivity disorders.

A retrospective cohort study of canid herpesvirus-1 seroprevalence and epidemiology in kennel dogs in Croatia

Canid alphaherpesvirus-1 (CaHV-1) is an endemic pathogen of dogs worldwide. CaHV-1 is often related to abortions, neonatal mortality, and the death of puppies. Since the first description of the virus in 1965, there has been no generally accepted method for diagnosing CaHV-1. Many authors used the virus neutralisation test (VNT) as a ‘reference standard’ due to its high specificity.Nasal, vaginal, preputial swabs and serum samples for this study were collected in the population of kennel dogs in Croatia. To determine the optimal VNT protocol, three modifications of the VNT were compared. These modifications were: VNT using native serum samples, VNT using thermally inactivated serum samples, and VNT using thermally inactivated serum samples with the addition of the complement. The correlation between the results of the VNT methods was significant (P < 0.001). Of all three modifications of VNT, the one using native serum samples was the one that increased VNT sensitivity. The overall seroprevalence of CaHV-1 was 32.02%. The PCR did not confirm the CaHV-1 presence in collected swabs.According to analysed anamnestic data, significant risk factors for CaHV-1 infection were: kennel size, attendance at the dog shows, hunt trials, kennel disinfection protocols, and mating. The oestrus cycle had no significant effect on seropositivity.The study results indicate that CaHV-1 spreads horizontally between dams living in kennels and in males during mating. Although there was no correlation between seropositivity and a history of reproductive disorders, significantly more stillborn puppies were recorded in seronegative dames (P < 0.01).

Immune interface of patients with reproductive loss in the North-West region of the Russian Federation

BACKGROUND: Currently, more than a hundred immunological biomarkers are known that are important for fertilization, implantation and placentation.&#x0D; AIM: The aim of this study was to assess the immunological profile for the main allo- and autoantibodies in women with reproductive loss and infertility in the North-West region of the Russian Federation.&#x0D; MATERIALS AND METHODS: This prospective cohort study was conducted at the stage of pregnancy planning in 325 women with reproductive disorders (spontaneous miscarriages, missed abortions, infertility, IVF failures) on the premises of the D.O. Ott Research Institute of Obstetrics, Gynecology, and Reproductive Medicine (St. Petersburg, Russia) from 2015 to 2022. The study assessed the immune interface of women with a history of reproductive disorders and matching loci of the HLA class II system (DQA1, DQB1, DRB1) among 100 married couples.&#x0D; RESULTS: In couples with reproductive disorders, coincidences of HLA class II allele (DQA1, DQB1, DRB1) loci were diagnosed in 14% (at 2 loci), 31% (at 3 loci), 14% (at 4 loci), 3% (at 5 loci) and 10% (at 6 loci) of couples. In 34.4% of women, we found an elevated number of spontaneously activated NK cells (CD107a). As many as 31.15% of women had a high level of antibodies to human chorionic gonadotropin. We found that 22.34% of women were carriers of at least one criteria antiphospholipid antibody, and 3.16% of women of two antiphospholipid antibodies. The diagnosis of antiphospholipid syndrome was not established in any patient. Non-criteria antiphospholipid antibodies were found in 6.15% of women. In 12.2% of women, we diagnosed a high level of antibodies to thyroperoxidase, while the average antibody value was 30.76 IU/ml. Only one patient had a high level of antisperm antibodies in the blood serum.&#x0D; CONCLUSIONS: In 59% of patients with a history of reproductive disorders, we identified some features of the immunological profile, which can lead to this pathology. Thus, 32% of women had a deviation in one immunological parameter, while 20% of patients had it in two parameters, and 7% in three or more parameters.

Serological Studies on Neosporosis and Risk Factors Assessment among Bovines of Haryana, India

Background: Bovine neosporosis causes reproductive disorders and abortion among bovines resulting in production and economic loss in meat and dairy industries. The current study was done to know about the status of neosporosis among bovines of Haryana State. Methods: A total of 165 serum samples from bovines of Haryana, having history of reproductive disorders or abortion were screened for brucellosis as well as neosporosis by ELISA. Result: All the samples were found as serologically negative for brucellosis, but seropositivity of 33.94% was observed for neosporosis by the monoscreen antibody ELISA. A significantly higher seropositivity was found among buffaloes (53.85%) than cows (21%), having an Odds ratio of 4.38. All the seropositive animals were associated with abortion with a prevalence rate of 36.36%, while none of male animals were diagnosed as seropositive. Bovines of age group 3-5 years were more seropositive. The bovines which had an abortion at third trimester of pregnancy were more seropositive (43.05%) than first (18.75%) and second trimester (38%) aborted bovines. The multiparous animals exhibited higher seropositivity (41.07%) in comparison to primiparous (23.81%) bovines. The present study confirms the presence of N. caninum antibodies in bovine population of Haryana State.

Presence of Chlamydia abortus in colostrum, milk and vaginal discharge samples of sheep

Background: The main transmission route of Chlamydia abortus is by ingesting the microorganism that has been eliminated in vaginal secretions, placental membranes or abortions that contaminate the environment and, possibly, through milk and colostrum. Elimination through vaginal secretions is well documented. However, there are no reports about isolation and identification of C. abortus in the colostrum or milk of infected sheep, so it is important to determine whether or not C. abortus may be present in these secretions, which are the only food of lambs. Objective: To detect C. abortus in colostrum, milk, and vaginal secretions of sheep with a history of reproductive disorders. Methods: Colostrum, milk, and vaginal exudates were collected from 66 sheep. The samples were inoculated in mouse fibroblast cell cultures and the presence of C. abortus determined by direct immunofluorescence. Results: 19 out of 66 colostrum samples (28.7%), 14 out of 66 milk samples (21.2%) and 17 out of 66 vaginal swabs (25.7%) were positive for C. abortus. The 50 samples positive for isolation and detected by immunofluorescence, together with 42 negative samples were subjected to qPCR to amplify a fragment of the ompA gene from C. abortus. Thirty-eight of the 92 samples processed by this technique were positive for C. abortus. Conclusion: The results demonstrated the presence of C. abortus in a high proportion in colostrum, milk and vaginal secretions of infected sheep. To the best of our knowledge, this is the first field study confirming the presence of C. abortus in colostrum, which shows that excretion of Chlamydia by lactogenesis could occur in the first hours after birth.

Infectious agents and reproductive disorders in non-human primates used as biomodels in animal experimentation

Reproductive disorders in non-human primates can be caused by a range of factors, in particular by infectious diseases. The literature review presented here aimed to identify the principal pathogens that can cause reproductive disorders in nonhuman primates, in particular, in the experimental models used for biomedical research. Although it is still largely unclear to what extent the pathogens as Coxiella burnetii, Brucella spp., Toxoplasma gondii, and Mycoplasma spp. can cause similar reproductive disorders in nonhuman primates, as observed in the human population and other animal group, we concluded that it would be strongly recommended to test for infection by these infectious agents all captive nonhuman primates with a history of reproductive disorders.

DNA polymorphisms and expression profile of immune and antioxidant genes as biomarkers for reproductive disorders tolerance/susceptibility in Baladi goat

The objective of this study was to explore single nucleotide polymorphisms (SNPs) and gene expression of immune and antioxidant markers associated with reproductive disorders in Baladi goats. A total of one hundred adults Baladi does were allocated into two equal-sized groups: normal reproductive performance and does have a history of reproductive disorders. DNA sequencing of PRLR (304-bp), LTF (904-bp), TLR2 (420-bp), TLR4 (335-bp), CLA-DRB3.2 (285-bp), SOD3 (735-bp), CAT (1526-bp), GPX4 (782-bp), and GST (690-bp) revealed SNPs associated with reproductive disorders tolerance/susceptibility in investigated does. Nonetheless, DNA sequencing of beta defensin (483-bp), CCL5 (840-bp), and ATOX1 (374-bp) genes elicited a monomorphic pattern. Levels of PRLR, LTF, TLR2, TLR4, CLA-DRB3.2, beta defensin, and CCL5 genes were significantly up-regulated in does affect with reproductive disorders than tolerant ones; while SOD3, CAT, GPX4, GST and ATOX1 genes pattern elicited an opposite trend. The results herein confirmed the potential significance of SNPs in immune and antioxidant genes as genetic markers for reproductive disorders tolerance/susceptibility in Baladi does. The Gene expression profile of investigated genes could be also used as proxy biomarkers for the prediction of the most susceptible risk time for disease occurrence and for building up an effective management protocol.

Characteristics of endometrial receptivity in women with different endometrial thickness

Aim: to evaluate the expression of estrogen (ER) and progesterone (PR) receptors in the endometrium during the "implantation window" in women with a history of reproductive dysfunction with different endometrial thickness.Materials and Methods. A prospective single-center cohort comparative study was conducted. The main group included patients with hypoplastic (&lt; 7 mm according to ultrasound data on preovulatory days) endometrium (n = 52), the comparison group consisted of women with normal endometrial thickness (n = 62) (women of both groups with reproductive dysfunctions of unclear cause), the control group included 16 healthy fertile women. An aspiration biopsy of the endometrium was performed on day 6–8 after ovulation with simultaneous collection of peripheral blood sample to measure level of estradiol (P). Endometrial biopsies were examined with histological and immunohistochemical (expression of ER, PR) methods.Results. All women had an ovulatory cycle, serum progesterone level ≥ 16.1 nmol/L (6–8 days after ovulation) and normoestrogenemia. E2/P was similar in all cohorts (p &gt; 0.05). Indicators of endometrial expression of ER, PR, similar to healthy women (n = 16), were found in 21 % (n = 11 out of 52) and 32 % (n = 20 out of 62) in women from main and comparison groups, respectively. The remaining 79 % (n = 41 out of 52) of women with "thin" endometrium (main group) and 68 % (n = 42 out of 62) of patients with normal endometrial thickness (comparison group) had ER and PR expression indices in the endometrial glands and stroma with significant differences (p &lt; 0.05) in all parameters (excepting PR in the endometrial stroma) from those of healthy women (control group), but without relevant differences between these cohorts (p &gt; 0.05) regardless of endometrial thickness.Conclusion. In women with a history of reproductive disorders, parameters of ER and PR expression similar to those in women without reproductive problems, were noted with a "thin" (&lt; 7 mm) endometrium by 50 % less often than those with normal thickness of the uterine mucosa. At the same time, the "thin" endometrium is not an absolute predictor of disorders of the hormone-receptor characteristics of the endometrium: in every fifth case, magnitude of endometrial expression ER, PR similar to healthy women were determined.

Neospora caninum infection and reproductive problems in dairy cows from Brazil: A case-control study.

Neosporosis, an infectious disease caused by the protozoan Neospora caninum, has been associated with economic losses in cattle rearing worldwide. However, previous studies have not presented any evidence regarding the association between serological status of neosporosis and alteration of the reproductive parameters. Thus, this study aimed to determine whether N. caninum is associated with reproductive disorders and to evaluate the possible risk factors of the infection. Blood samples from 202 dairy cows, 51 with a history of reproductive disorders (case group) and 151 without (control group), were collected from different farms in Brazil. Epidemiological questionnaires were conducted with all the farmers. Serum samples were subjected to an indirect fluorescent antibody test to detect antibodies against the parasite. In total, 28.22% (57/202) of the cows were seropositive: 47.06% (24/51) from the case group and 21.85% (33/151) from the control group. By logistic regression, cows aged ≥48months and cows with history of abortion were 4.8 (95% confidence interval [CI]=1.91-12.05; p=0.001) and 2.3 (95% CI=1.06-5.1; p=0.034) times more likely to be seropositive, respectively. Furthermore, our results show an association between N. caninum seropositivity and abortion in dairy cows from Brazil with poor management conditions and N. caninum seropositivity risk factors for reproductive disorders.

SERO-MOLECULAR INVESTIGATION OF COXIELLA BURNETII INFECTION IN DOMESTIC RUMINANTS AND HUMANS AND ASSOCIATED RISK FACTORS BASED ON ‘ONE HEALTH’ PERSPECTIVES IN BANGLADESH

Background: Q-fever is an important zoonotic disease caused by the bacterium Coxiella burnetii with wide host range of mammals, birds and arthropods worldwide. The prevalence of C. burnetii infection has been reported in domestic ruminants in Bangladesh with no attention on the ‘One Health’ approach for the epidemiological investigation associated with risk factors for prevention of the disease. Objective: This study aimed to investigate the sero-molecular prevalence of Coxiella burnetii infection and associated risk factors in domestic ruminants and humans based on ‘One Health’ approach in Bangladesh Materials and Methods: This study on C. burnetii infection was conducted in cattle, goats and humans of the four randomly selected districts (Kurigram, Sirajgonj, Pabna and Mymensingh) in Bangladesh during the period from 2018 to 2021. A total of 162 and 172 serum samples respectively from cattle and goats with the reproductive disorders, 159 serum samples from human patients with pyrexia of unknown origin lasting over a period of three weeks and who were in close contact with animal, 119 milk samples from cows with reproductive disorders and 6 aborted materials from goat were collected from these selected districts for the detection of C. burnetii antibody by ELISA and DNA by PCR assay. The research was performed with the collaboration of OIE reference laboratory for Q fever, Friedrich-Loeffler-Institute, Germany. Results: The overall sero-prevalence of C. burnetii infection by ELISA was 3.01% in cattle, 7.6 % in goats and 0.63% in humans. The highest sero-positivity was recorded in cattle in Mymensingh (4.5%) in comparison to Pabna (2.8%), Kurigram (2.5%) and Sirajgonj (2.4%) districts. Risk factors associated with higher sero-positivity of C. burnetii infection was recorded in cattle with  5 years (4.4%) than 3 to 5 years (2.1%), indigenous (4.2%) than cross-bred (2.6%), history of retention of placenta (5.6%) than abortion (2.3%) and natural service (5.9%) than artificial insemination (2.3%). The sero-positivity of C. burnetii infection in goat showed highest in Kurigram (10.0%), followed by Sirajgonj (9.5%) and Mymensingh (9.3%) with no positive reactors in Pabna district. Risk factors associated with sero-positivity of C. burnetii infection in goats showed higher in &gt; 2 years (14.1%) than up to 2 years (2.1%) age group and higher with the history of abortion (10.0%) than retention of placenta (4.2%). Risk factors associated with positivity of C. burnetii infection detected in milk samples of cows by ELISA showed an overall 21.85% positivity associated with history of reproductive disorders, with highest positivity in anestrus (33.3%), followed by retention of placenta (24.4%), abortion (21.7%) and lowest with repeat breeding (8.3%). In addition, higher positivity was recorded in cross-bred (24.2%) than in indigenous (12.5%) cattle. In case of humans, only one human patient (0.63%) had positive for IgG phase-II ELISA. C. burnetii DNA was detected in two sero-positive milk samples but all of the intermediate positive milk samples by ELISA were negative by PCR assay and even none of the aborted material of goat was positive in PCR and cell culture. Conclusions: This study recorded the prevalence and risk factors associated with the zoonotic C. burnetii infection in both the domestic ruminants and humans but still it is unrecognized and underestimated in both human and animal health and research in Bangladesh. The findings of this study support the further research on C. burnetii in both human and veterinary medicine under ‘One Health’ approach particularly targeting epidemiology on the agent, host and environment for the control and prevention of the disease in Bangladesh. Keywords: C. burnetii, Ruminants, Humans, ELISA, PCR, Sera, Milk, Prevalence, Risk factors, ‘One Health’ approach

Abortion Due to Neospora caninum in Dairy Cattle in Southern Santa Catarina State, Brazil

Background: Neosporosis is a cosmopolitan disease known as the main infectious cause of abortion in cattle, reported in several states in Brazil. The transplacental transmission in cattle is responsible for perpetuating the disease in the herd. In the state of Santa Catarina, previous studies on this protozoan in cattle are mostly serological surveys. To increase information about this reproductive disorder, this work describes the diagnosis of abortions due to Neospora caninum in dairy cattle from state of Santa Catarina and the follow-up for 4 years in a farm affected by neosporosis.Case: From 2015 to 2019, necropsy was performed on 10 aborted bovine fetuses, between fifth and eighth month of pregnancy, with 1 fetus aborted in 2015, 3 in 2016, 2 in 2017 and, 4 in 2019, all originating from the same dairy property in the southern region of state of Santa Catarina. No macroscopic lesions were found. Histopathology revealed mild to moderate multifocal mononuclear necrotizing encephalitis in 5 fetuses, and multifocal mild mononuclear myositis and myocarditis in 4 and 3 fetuses, respectively. Polymerase Chain Reaction (PCR) using primers based on the Nc-5 gene was positive for N. caninum in five fetuses. Three visits were performed in the farm for epidemiological evaluation and blood samples collection for IgG antibodies anti-N. caninum (IFAT). The total herd was 170 Jersey, Holstein and crossbred cattle, raised in a semi-confined system with mechanical milking system. Since 2014, there has been a significant increase in abortions cases, approximately 20 cows had abortions; in the year 2015, approximately 10; in 2016, less than 5; in 2017, 4 cows aborted; in 2018, 11 abortions and, in 2019, there were 4 abortions. An increase in the rate of return to estrus was also reported, and both primiparous and multiparous cows had reproductive disorders. Abortions were recorded throughout the year and occurred predominantly between the fourth and sixth month of pregnancy. In 2016, an Indirect Immunofluorescence Reaction (IFAT) was performed on sera from 26 cows (13 with a history of reproductive disorders and another 13 without disorders). Of these, 50.0% (13/26) were seropositive, with titers ranging from 100 to 1600 (cutoff ≥100). The Enzyme Immunoassay (ELISA) of the entire herd was carried out in 2017, with 26.54% (43/162) of seropositivity, 8.02% (13/162) suspect, and 30.4% (17/56) had reproductive disorders. It was observed that only animals born on the property were used for replacement, there was less possibility of direct contact between dogs on the property with milking facilities, placental and fetal remains, properly disposing of these, incinerating or burying. Thirty animals were discarded from the property, 25 of which had reproductive disorders. All young female daughters of seropositive cows remained on the property for replacement. During the evaluation period, all dogs were eliminated.Discussion: In this study, the diagnosis of neosporosis was made through epidemiology, histopathological lesions characterized by mononuclear encephalitis, myocarditis and myositis, and detection of the agent by PCR, associated with serological techniques. The lesions observed are indicative of N. caninum infection and are compatible with lesions observed in other studies. Serological screening is important to complement the diagnosis of abortion by N. caninum and to help control the agent in herds. From this report it is concluded that N. caninum is an important agent of reproductive disorders in cattle in the southern region of Santa Catarina, the different serological analyzes showed a good screening index for the inclusion of control strategies. In addition, the monitoring of reproductive rates of affected properties becomes necessary over the years, allowing better observation of control strategies.Keywords: reproductive disorder, fetal death, protozoan, parasitology.Descritores: distúrbio reprodutivo, morte fetal, protozoário, parasitologia.

Is Bluetongue Virus a Risk Factor for Reproductive Failure in Tropical Hair Sheep in Brazil?

Background: Bluetongue is a vector-borne viral disease transmitted by midges from the genus Culicoides. The disease can infect most of the ruminant and camelid species, but the severe disease is most often seen in european wool and mutton sheep breeds. In this sense, there is a gap in the knowledge on BTV infection in hair sheep breeds from tropical zones. Thus, this study aimed at establishing whether exposure to BTV is a risk factor for reproductive failure in Santa Inês ewes, a hair sheep breed, reared under tropical conditions in Brazil.Materials, Methods &amp; Results: A retrospective cross-sectional study was carried out in sheep farms in São Paulo state, Brazil, after the rainy season. Serum samples from 110 Santa Inês ewes with a history of reproductive disorders, in the last 6 months, which were included: abortion, premature birth, stillbirth, retention of placenta, infertility, estrus repetition, fetal malformation, weak lamb birth and neonatal death were collected. The presence of antibodies against BTV was assessed by agar gel immunodiffusion method (AGID). Serology to the infectious agents Brucela ovis, Lepstopira spp., Toxoplasma gondii, Neospora caninum and Campylobacter sp. were also assessed. Bivariate associations between the outcome and individual explanatory variables were assessed using the Fisher's exact test. Abortion was the most common reproductive disorder (53%; 74/139) observed, followed by estrus repetition (12%; 17/139) and infertility (11%; 15/139). Other disorders related to the conceptus totaled nearly one fourth of the reported disorders. A total of 20% (22/110) of the ewes were seropositive to BTV. A higher frequency of BTV seropositive than BTV seronegative ewes with a history of abortion was found. Also, abortion with seroreactivity to BTV was tested for prevalence ratio that showed 1.38 [95% CI 1.10-1.74; P = 0.030]. With regards to the abortion involvement of other infectious diseases associated with the seropositive ewes to BTV, more than a half of ewes (53%; 10/19) were solely seropositive for BTV.Discussion: In the current study, it was detected 20% (22/110) of seropositive ewes to BTV. These findings demonstrated that even though the BTV has been considered endemic in tropical countries such as Brazil, there are regions or microclimates in which the virus cannot be present or in varied prevalence. The history of abortion was identified as the potential factor associated with BTV seropositivity in Santa Ines ewes. Equally, the differential diagnosis for other infectious agents related to abortion demonstrated the unique presence of antibodies against BTV in more than half of all cases. Other studies with native sheep flocks in Iran and Nepal also demonstrated a strong positive correlation between abortion history and seropositivity for BTV. Thus, it is possible that in other continents of the world, under tropical conditions, the virus does not behave the same asymptomatic infection such as have been reported for native sheep breeds in Africa. In conclusion, it was demonstrated that one-fifth animals were positive for antibodies against BTV clearly implying the viral spreading in the local hair sheep flocks. These findings highlight the importance of surveillance related to BTV in endemic areas. Therefore, it is recommended to strengthen the surveillance system for BTV within Brazil and to educate farmers about the management and control of this disease.

Epidemiology and Associated Risk Factors for Brucellosis in Small Ruminants Kept at Institutional Livestock Farms in Punjab, Pakistan.

Brucellosis is reportedly endemic in ruminants in Pakistan. Both Brucella abortus and B. melitensis infections have been decumented in domestic animals and humans in the country. This study aimed to identify the burden of anti-Brucella antibodies in small ruminants as well as associated potential risk factors with its occurrence at nine institutional livestock farms in Punjab, Pakistan. The sera collected from equal number of sheep and goats (500 from each species) were screened by indirect-ELISA for anti-smooth-Brucella antibodies followed by a serial detection by real-time PCR. Overall, 5.1% (51/1000) seropositivity was registered corresponding to 5% (25/500) prevalence in goats and 5.2% (26/500) in sheep. Brucella-DNA could not be detected in any of the tested sera by real-time PCR. Multiple logistic regression model indicated that farm location (OR 34.05), >4 years of age (OR 2.88), with history of reproductive disorders (OR 2.69), and with BCS of ≤ 3 (OR 12.37) were more likely to test positive for brucellosis at these farms. A routine screening, stringent biosecurity, and quarantine measures are warranted for monitoring and eradication of the infection. Similarly, isolation and molecular investigation of the etiologic agent(s) are needed to understand the relationship of epidemiology and out-breaks of brucellosis in the country.

Почечная дисфункция при преэклампсии: прогнозирование и дифференциальный диагноз. Часть 2

Study Objective: To assess the prognostic value, sensitivity, and specificity of biomarkers of acute renal injury (ARI), and of angiogenic factors, in the development of pre-eclampsia (PE) in the second trimester of pregnancy, and determine their diagnostic significance for PE, as well as to compare obstetric and perinatal outcomes experienced by patients. Study Design: This was a comparative, group, prospective and retrospective study. Materials and Methods: The patient groups studied in the first, second, and third phases are described in Part 1 of this article. Based on the distinguishing diagnostic criterion and the outcomes of pregnancy, the 138 patients were divided into three groups. Group I (main) consisted of 66 (47.8%) patients who developed hypertensive disorders during pregnancy, 30 (21.7%) of whom were included in subgroup 1 (CKD) and 36 (26.1%) in subgroup 2 (no CKD). Group II (comparison) was made up of 32 patients with CKD who did not have hypertensive disorders. Group III (control) comprised 40 women with normal pregnancies and no history of reproductive disorders. The fourth phase involved a retrospective analysis of biomarker levels and assessment of their prognostic value for the development of PE. All participants underwent clinical and laboratory examinations and had measurements taken for sFlt-1, PlGF, S-endoglin, cystatin C, uKIM-1, podocalyxin, and α1- and β2-microglobulins. Obstetric and perinatal outcomes were traced. Study Results: Levels of the following parameters had the greatest prognostic value in the second trimester of pregnancy: PlGF (94%), sFlt-1 (92%), sFlt-1/PlGF (94%), ΔPlGF (93.3%), ΔsFlt-1 (92%), ΔsFlt-1/PlGF (94%), S-endoglin (94%), and sNGAL (94%). The sensitivity of the sFlt-1/PlGF ratio at this stage of pregnancy was 89.1%, making this parameter a promising predictive marker of PE. Logistic regression analysis showed that it is most reasonable to measure sFlt-1/PlGF and sNGAL levels in the first trimester; and in the second trimester, ΔsFlt-1/PlGF (between the first and second trimesters) and sFlt-1/PlGF as well as levels of PlGF, S-endoglin, uKIM-1, urinary podocalyxin, and sNGAL. In both subgroups of patients with PE, there were critical hemodynamic disruptions in the fetal-placental-maternal system: six (20%) and eight (22.2%) cases in subgroups 1 and 2, respectively. Pre-term delivery in patients with pre-existing intrauterine growth retardation led to the necessity of putting 22 (73.3%) newborns from the first subgroup and seven (19.4%) newborns from the second subgroup on mechanical ventilation (p&lt;0.0001). Conclusion: Some weeks before the clinical onset of PE, at weeks 16-24 of pregnancy, patients develop placental dysfunction, decreases or slight increases in PlGF levels, and elevation of sFlt-1 levels, reflecting an imbalance between pro-angiogenic and anti-angiogenic factors; there is also an increase in the levels of markers of ARI (sNGAL, uKIM-1, β2- and α1-microglobulins, and urinary podocalyxin). The clinical information obtained about perinatal outcomes indirectly confirms the role of the cascade of pathogenic events in chronic placental insufficiency, in the development of generalized endothelial dysfunction. Keywords: pre-eclampsia, predictive diagnosis, chronic kidney disease, sFlt-1, PlGF, S-endoglin, cystatin C, KIM-1, podocalyxin, α1-microglobulin, β2-microglobulin.

Epidemiology of Sub-Clinical Mastitis in Dairy Cows in Urban Areas of Chittagong, Bangladesh

Mastitis is considered to be the most devastating condition for the dairy farms of low and medium income countries due to its alarming impact on production and worldwide has been recognized as one of the most economically significant infectious diseases affecting the welfare of dairy animal. Chittagong is one of the dairy intensive regions of Bangladesh but comprehensive epidemiological studies addressing prevalence of and risk factors for subclinical mastitis (SCM) are scant. Therefore we aimed to approximate the prevalence of SCM in dairy cows along with the associated risk factors in dairy cows of Chittagong Metropolitan Area (CMA), Bangladesh. We included 114 cross breed (Holstein × Local) lactating cows, raised at 6 dairy farms in CMA between February to November, 2015 for this study. California Mastitis Test (CMT) was used to assess SCM at animal level. The overall prevalence of mastitis was 34.2% among the tested cows. Floor type, source of replacement cows, history of previous reproductive disorder, stage of lactation and cleanliness of floor was found significantly associated with SCM in univariate analysis. In random effect multiple logistic regression model, cemented floor was found to increase the odds of SCM by 5.03 times than that of brick floor. Similarly, cows with history of reproductive disorders had more risk of having SCM than that of cows without a history of reproductive disorders. Since the SCM is prevalent in the study area, the intervention strategy should focus on causal agent, improved management, frequent monitoring of SCM in milking cows with CMT and use of teat disinfectants; with minimal or no treatment with antimicrobial agents.

Features of the hormone-receptor interaction in the endometrium during ovulatory menstrual cycle in women with reproductive failure

A comparative analysis of the levels of estradiol and progesterone in the blood, immunohistochemical parameters of estrogen receptors and progesterone in endometrium in women with a history of reproductive disorders is presented. It was found that all women had an ovulatory ovarian cycle, levels of estradiol and progesterone in the blood were within the reference values. In women with reproductive failures in the history (n=107), four types of hormone-receptor response in the endometrium were identified. In 46 (43%) women, the first (normoreceptor) type of endometrial response was detected, without significant differences from the control group (n=15) corresponding to the middle stage of the secretion phase. Hyperreceptor (hyper-estrogen-progesterone-receptor, hyper-estrogen-receptor, hyper-progesterone-receptor) types have been identified in 61 patients (57%) with reproductive dysfunctions. The endometrium corresponded to the mid secretory phase was detected in 47 (44%), inadequate secretory phase of the endometrium - in 60 (56%) women with reproductive failures in the anamnesis. All women in the control group had a full secretory change in the endometrium. In general, more than half (61 (57%)) of women with reproductive failures in the history with the ovulatory menstrual cycle with normal values of the level of progesterone in the blood showed signs of a decreased endometrial receptivity status. This indicates that the ovulatory level of progesterone in the blood is not an unconditional predictor of full secretory transformations of the endometrium.