Parental positive affect and negative affect in same- and different-sex parent families: no associations with parental gender and caregiving role.

Positive and negative parental affect influence developing parent-child attachment relationships, especially during infancy as well as children's social-emotional, academic, and behavioral functioning later in life. Increasingly, because both mothers and fathers can play central caregiving roles, the parenting qualities of both parents demand consideration. Therefore, this study investigated whether parental gender and caregiving role were associated with mothers' and fathers' positive affect and negative affect during interactions with their 4-month-old firstborn infant, while determining whether parenting stress, infant temperament, having a singleton/twin, and living in the Netherlands, France, or the United Kingdom were related to parental positive affect and negative affect. In all, 135 different-sex, same-sex male, and same-sex female couples (113 fathers and 157 mothers, comprising 147 primary, and 123 secondary caregivers) who conceived through artificial reproductive techniques were studied. The couples were videorecorded at home while in feeding, cleaning, and playing contexts to assess the levels of positive and negative parental affect. In addition, the couples completed questionnaires about their caregiving role, parenting stress, and the infants' temperament. Mixed linear models indicated that the levels of positive and negative parental affect toward the infant in all contexts were not related to parental gender, caregiving role, the interaction between parental gender and caregiving role, parenting stress, infant temperament, or singleton/twin status. However, the target parental behaviors were related to the country of origin, suggesting differences among Dutch, French, and British parents. Overall, we found no evidence that gender or caregiving roles were associated with the levels of positive and negative affect shown by the parents.

Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii.

Neosporosis and toxoplasmosis are major causes of abortion in livestock worldwide, leading to substantial economic losses. Detection tools are fundamental to the diagnosis and management of those diseases. Current immunohistochemistry (IHC) tests, using sera raised against whole parasite lysates, have not been able to distinguish between Toxoplasma gondii and Neospora caninum. We used T. gondii and N. caninum recombinant proteins, expressed in Escherichia coli and purified using insoluble conditions, to produce specific polyclonal rabbit antisera. We aimed to develop species-specific sera that could be used in IHC on formalin-fixed, paraffin-embedded (FFPE) tissue sections to improve the diagnosis of ruminant abortions caused by protozoa. Two polyclonal rabbit sera, raised against recombinant proteins, anti-Neospora-rNcSRS2 and anti-Toxoplasma-rTgSRS2, had specificity for the parasite they were raised against. We tested the specificity for each polyclonal serum using FFPE tissue sections known to be infected with T. gondii and N. caninum. The anti-Neospora-rNcSRS2 serum labeled specifically only N. caninum-infected tissue blocks, and the anti-Toxoplasma-rTgSRS2 serum was specific to only T. gondii-infected tissues. Moreover, tissues from 52 cattle and 19 sheep previously diagnosed by lesion profiles were tested using IHC with our polyclonal sera and PCR. The overall agreement between IHC and PCR was 90.1% for both polyclonal anti-rNcSRS2 and anti-rTgSRS2 sera. The polyclonal antisera were specific and allowed visual confirmation of protozoan parasites by IHC, but they were not as sensitive as PCR testing.

Donor sibling relations among adult offspring conceived via insemination by lesbian parents.

STUDY QUESTION: How do adult offspring in planned lesbian-parent families feel about and relate to their donor (half) sibling(s) (DS)? SUMMARY ANSWER: A majority of offspring had found DS and maintained good ongoing relationships, and all offspring (regardless of whether a DS had been identified) were satisfied with their knowledge of and contact level with the DS. WHAT IS KNOWN ALREADY: The first generation of donor insemination offspring of intended lesbian-parent families is now in their 30s. Coincident with this is an increased use of DNA testing and genetic ancestry websites, facilitating the discovery of donor siblings from a common sperm donor. Few studies of offspring and their DS include sexual minority parent (SMP) families, and only sparse data separately analyze the offspring of SMP families or extend the analyses to established adult offspring. STUDY DESIGN, SIZE, DURATION: This cohort study included 75 adult offspring, longitudinally followed since conception in lesbian-parent families. Quantitative analyses were performed from online surveys of the offspring in the seventh wave of the 36-year study, with a 90% family retention rate. The data were collected from March 2021 to November 2022. PARTICIPANTS/MATERIALS, SETTING, METHODS: Participants were 30- to 33-year-old donor insemination offspring whose lesbian parents enrolled in a US prospective longitudinal study when these offspring were conceived. Offspring who knew of a DS were asked about their numbers found, characteristics or motivations for meeting, DS terminology, relationship quality and maintenance, and impact of the DS contact on others. All offspring (with or without known DS) were asked about the importance of knowing if they have DS and their terminology, satisfaction with information about DS, and feelings about future contact. MAIN RESULTS AND THE ROLE OF CHANCE: Of offspring, 53% (n = 40) had found DS in modest numbers, via a DS or sperm bank registry in 45% of cases, and most of these offspring had made contact. The offspring had their meeting motivations fulfilled, viewed the DS as acquaintances more often than siblings or friends, and maintained good relationships via meetings, social media, and cell phone communication. They disclosed their DS meetings to most relatives with neutral impact. The offspring, whether with known or unknown DS, felt neutral about the importance of knowing if they had DS, were satisfied with what they knew (or did not know) of the DS, and were satisfied with their current level of DS contact. This study is the largest, longest-running longitudinal study of intended lesbian-parent families and their offspring, and due to its prospective nature, is not biased by over-sampling offspring who were already satisfied with their DS. LIMITATIONS, REASONS FOR CAUTION: The sample was from the USA, and mostly White, highly educated individuals, not representative of the diversity of donor insemination offspring of lesbian-parent families. WIDER IMPLICATIONS OF THE FINDINGS: While about half of the offspring found out about DS, the other half did not. Regardless of knowing of a DS, these adult offspring of lesbian parents were satisfied with their level of DS contact. Early disclosure and identity formation about being donor-conceived in a lesbian-parent family may distinguish these study participants from donor insemination offspring and adoptees in the general population, who may be more compelled to seek genetic relatives. The study participants who sought DS mostly found a modest number of them, in contrast to reports in studies that have found large numbers of DS. This may be because one-third of study offspring had donors known to the families since conception, who may have been less likely to participate in commercial sperm banking or internet donation sites, where quotas are difficult to enforce or nonexistent. The study results have implications for anyone considering gamete donation, gamete donors, donor-conceived offspring, and/or gamete banks, as well as the medical and public policy professionals who advise them. STUDY FUNDING/COMPETING INTEREST(S): No funding was provided for this project. The authors have no competing interests. TRIAL REGISTRATION NUMBER: N/A.

Sperm donor relations among adult offspring conceived via insemination by lesbian parents.

AIM: The present study examined how adult offspring of lesbian parents relate to their anonymous, open-identity, or known donors. DESIGN: An online survey of 75 donor-conceived offspring of lesbian parents, aged 30-33 years, participating in Wave 7 of a U.S. 36-year longitudinal study of planned lesbian-parent families was conducted. Offspring were asked about donor type, motivations for contacting the donor, terminology for the donor, relationship quality, means of relationship maintenance, impact of donor contact on offspring's other family members, and their feelings about the donor. RESULTS: Twenty offspring with anonymous donors and 15 with open-identity donors whom they had not contacted felt comfortable not knowing their donors. Forty offspring knew their donors - anonymous, contacted through an online registry (n = 7), open-identity, contacted (n = 9), or known since childhood (n = 24). Offspring who had contacted their donor since age 18 had their motivations fulfilled after contact, got along well with him, did not view him as a relative, and had told most family members about their contact, without detriment. Whether the donor was unknown or known at this stage of their lives, most offspring were satisfied with their contact level. CONCLUSION: This cohort of donor-conceived offspring of lesbian parents was among the first to reach adulthood during a time of technological advances in DNA testing, giving access to anonymous donors via online registries. The results inform donors, families, mental health providers, medical providers and public policymakers, on whether, how, and to what degree donor-conceived offspring optimally make donor contact.

Gay Community Stress in Sexual Minority Men and Women: A Validation Study in the Netherlands.

Intraminority gay community stress theory posits that social stressors within sexual minority communities of men may be risk factors for mental health problems in gay and bisexual men. The recently developed 20-item Gay Community Stress Scale (GCSS) is a valid and reliable measure of gay community stress, but was not yet validated in the Netherlands. This study developed a Dutch-translated version of the GCSS and validated this scale in sexual minority men and sexual minority women, as it was hypothesized that sexual minority women may also experience intraminority stress. Exploratory and confirmatory factor analyses were subsequently performed in independent samples of men and women, and produced a 16-item GCSS for men and a 12-item GCSS for women. The four-factor structure of the original GCSS was replicated in men and women, and encouraging support for discriminant and concurrent validity of the GCSS was found in both men and women. The total scale and subscales were internally consistent in men (α and ω ≥ .87) and in women (α and ω ≥ .78). The Dutch-translated GCSS seems to offer a valid and reliable way to assess intraminority stress in Dutch-speaking sexual minority men and sexual minority women, although further validation is warranted.

Long-Term Effects of Family Resilience on the Subjective Well-Being of Offspring in the National Longitudinal Lesbian Family Study.

The current study used a family resilience approach to investigate why some offspring of sexual minority parents thrive despite homophobic stigmatization while others do not. Specifically, the study explored the role of two specific family functioning mechanisms (i.e., during adolescence, disclosure of offspring's personal life to their parents, and family compatibility) in the association between experienced homophobic stigmatization at age 17 and subjective well-being at age 25, among 71 offspring (37 females and 34 males, all cisgender) of the National Longitudinal Lesbian Family Study (NLLFS). The results showed that, overall, the offspring reported healthy subjective well-being as emerging adults. However, among NLLFS offspring with less family compatibility as adolescents, homophobic stigmatization was related to higher scores on negative affect when they were emerging adults. Psychological counseling that supports adolescent-parent communication may help prevent the negative effect of homophobic stigmatization on the subjective well-being of offspring with sexual minority parents.

Mental Health Disparities Between Lesbian Mothers and Heterosexual Parents: the Mediating Role of Positivity.

Introduction: Previous evidence has shown better mental health outcomes for lesbian mothers, in comparison with heterosexual parents. The present study explored the mediating role of positivity (i.e., the tendency to view life with a positive outlook) in determining these mental health differences. Methods: Seventy-two Israeli lesbian mothers by donor insemination (from 36 families) were compared with 72 Israeli heterosexual parents by assisted reproduction (without donated gametes; from 36 families) on positivity, life satisfaction, and depressive symptomatology. All parents were cisgender and had at least one child born through assisted reproduction, aged 3-10 years. Data were collected between December 2019 and February 2021. Results: Mediational analyses indicated that lesbian mothers reported greater positivity, which was, in turn, associated with greater life satisfaction and less depressive symptomatology. Conclusions: The results pinpoint the possible protective and strengthening role of positivity for lesbian mothers in the pronatalist and familistic environment of Israel. The research contributes to the literature on the mental health of parents with diverse sexual orientations living outside of the USA and Europe. Policy Implications Policymakers and mental health professional should be knowledgeable about the importance of improving positivity as a strategy for reducing depressive symptomatology and promoting life satisfaction among lesbian mothers. Moreover, policy makers and legislators are called to ease the access of sexual minority individuals to parenthood, as in this population parenthood associates with heightened mental health outcomes.

Assisted Conception Socialization Self-Efficacy Among Israeli Lesbian, Gay, and Heterosexual Parent Families and its Association with Child Externalizing Problems.

This questionnaire-based study compared 36 Israeli lesbian mother families (n = 72 lesbian mothers) formed by donor insemination, 39 Israeli gay father families (n = 78 gay fathers) formed by gestational surrogacy, and 36 Israeli heterosexual parent families (n = 72 heterosexual parents) formed by assisted reproduction (without donated gametes), all with a target child aged 3-10 years. The families were examined for parents' assisted conception socialization self-efficacy, depression, negative and positive affect, life satisfaction, positivity, resilience, social support, and child externalizing problems. Multiple factors associated with child externalizing problems were also examined. Multilevel modeling analyses indicated that parents' assisted conception socialization self-efficacy did not differ between family groups; however, lesbian mothers and gay fathers reported fewer child externalizing problems and greater social support, relative to heterosexual parents. Also, lesbian mothers-but not gay fathers-reported lower levels of depression, greater life satisfaction, and more positivity than did heterosexual parents. Finally, irrespective of family type, greater assisted conception socialization self-efficacy was associated with fewer parent-reported child externalizing problems. Findings are interpreted in light of the cultural socialization framework and Israel's familistic and pronatalist environment. Implications for health professionals, educators, and policymakers working with diverse family forms are discussed.

The desire for more children among Israeli lesbian, gay, and heterosexual couples who became parents through assisted reproduction.

The present online questionnaire-based study compared 72 lesbian mothers by donor insemination (within 36 families), 78 gay fathers by gestational surrogacy (within 39 families), and 72 heterosexual parents by assisted reproduction (without donated eggs and/or sperm; within 36 families), on their desire to have more children and their ideal number of children. All participants self-identified as cisgender, lived in Israel, and had a mean number of two children. The study also explored the predictors of participants' desire for more children. Multilevel modeling analyses indicated that, when participants' actual number of children was controlled for, lesbian and gay parents did not differ from heterosexual parents on their desire for more children and their ideal number of children. Parents' younger age, greater positivity, and lower attribution of responsibility to children for misbehavior, in addition to a lower discrepancy between their actual and ideal number of children, were associated with greater desire for more children. The results suggest that lesbian and gay parenthood may mark an entrance to the Israeli familistic "mainstream," which, in turn, may decrease differences in parenthood aspirations as a function of sexual orientation. The study contributes to the scant literature on the desire for more than one child among parents in diverse family configurations. It further provides family researchers and policy makers with vital information on the most influential factors associated with the desire for more children among lesbian, gay, and heterosexual parents. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Helicopter parenting, emotional avoidant coping, mental health, and homophobic stigmatization among emerging adult offspring of lesbian parents.

Helicopter parents are highly involved parents who hover over and around their child, applying developmentally inappropriate levels of control and tangible assistance. Previous research with different-sex parent families indicates that helicopter parenting is particularly problematic in emerging adulthood as it may indirectly affect the offspring's mental health through their use of emotional avoidant coping. Knowledge is lacking, however, on the antecedents and consequences of helicopter parenting in lesbian-parent families. The present longitudinal, questionnaire-based study investigated the effect of homophobic stigmatization in adolescence on mental health via helicopter parenting and emotional avoidant coping among 76 (37 females and 39 males) National Longitudinal Lesbian Family Study 25-year-old offspring of lesbian parents. All participants were cisgender, born in the USA, and conceived through donor sperm, with the majority being White, heterosexual, highly educated, and no longer living with their parents. Parents who reported that their offspring experienced homophobic stigmatization in adolescence were likely to enact higher helicopter parenting in emerging adulthood. Then, higher scores on helicopter parenting were associated with offspring's greater use of emotional avoidant coping, which in turn negatively affected the mental health of emerging adult offspring. Discussed in light of Bowen's family differentiation theory, the results suggest that clinicians should examine helicopter parenting in the context of lesbian parents' developmental history and potential tendency to project their own concerns about safety onto their child in order to reduce the distress of experienced homophobic stigmatization. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Behavioral Outcomes of Children with Same-Sex Parents in The Netherlands.

Same-sex parents face substantial stressors due to their sexual orientation, such as experiences of prejudice and prohibitive legal environments. This added stress is likely to lead to reduced physical and mental health in same-sex parents that, in turn, may translate into problematic behavioral outcomes in their children. To date, there are only a few nationally representative studies that investigate the well-being of children with same-sex parents. The current study takes a closer look at children's behavioral outcomes, reported by a parent, using an adapted version of the emotional, conduct, hyperactivity, pro-social, and peer problems subscales of the Strengths and Difficulties Questionnaire (SDQ). We take advantage of unique data from the Netherlands based on a probability sample from population registers, whereby findings can be inferred to same-sex and different-sex parent households with parents between the ages of 30 and 65, and with children between the ages of 6 and 16 years (62 children with same-sex, and 72 children with different-sex parents). The findings obtained by coarsened exact matching suggest no significant disadvantages for children with same-sex parents compared to different-sex parents. We contextualize these findings in their wider cultural context, and recommend a renewed focus in future research away from deficit-driven comparisons.

Unmet support needs in donor sperm treatment: consequences for parents and their donor-children.

RESEARCH QUESTION: Are unmet needs for psychosocial counselling, peer support and friends/family support in parents directly and/or indirectly related to the mental health of parents and their donor-children? DESIGN: A cross-sectional sample of 214 parents participated in this quantitative study via an online questionnaire. The sample comprised mothers and fathers in a heterosexual relationship (n = 85), mothers in a lesbian relationship (n = 67) and single mothers (n = 62). Parents were recruited via three Dutch fertility clinics and four network organizations. Unmet support needs were measured with an adapted version of the Unmet Needs for Parenting Support questionnaire, changing the original items into items about donor conception. The items were derived from a qualitative study and checked by experts in donor conception. The parents' mental health was measured with the Adult Self Report and the donor-children's mental health with the Child Behaviour Checklist. A multigroup mediation analysis was conducted to explore relationships between parents' unmet support needs and their child's mental health, with the parents' mental health as a possible mediator. RESULTS: There were no direct relations between parents' unmet support needs and the mental health of donor-children. Unmet needs for psychosocial counselling, peer support and friends/family support for parents and children's mental health were indirectly related through the mental health of the parents: 0.074 (CI 95%  = 0.013-0.136; P = 0.017), 0.085 (CI 95% = 0.018-0.151; P = 0.036) and 0.063 (CI 95% = 0.019-0.106; P = 0.013), respectively. CONCLUSIONS: We recommend that fertility clinics, network organizations and authorities for infertility counsellors make their support available to parents for extended periods after their treatment. Further qualitative studies are necessary to assess how to relieve unmet support needs during donor sperm treatment.

Minority Stress and Suicidal Ideation and Suicide Attempts Among LGBT Adolescents and Young Adults: A Meta-Analysis.

Purpose: This meta-analytic study examined associations between minority stressors and suicidal ideation and suicide attempts among LGBT adolescents and young adults (aged 12-25 years). Methods: Identified studies were screened using the inclusion and exclusion criteria. Studies had to include an association between a minority stressor and a suicidality outcome and were categorized into 10 meta-analyses. Overall effect sizes were calculated using three-level meta-analyses. In addition, moderation by sampling strategy was examined. Results: A total of 44 studies were included. Overall, LGBT bias-based victimization, general victimization, bullying, and negative family treatment were significantly associated with suicidal ideation and/or suicide attempts. Associations of discrimination and internalized homophobia and transphobia with suicidal ideation and/or suicide attempts were not significant. No moderation effects were found for sampling strategy. Conclusion: Although overall effect sizes were small, our meta-analytic study shows a clear link between various types of minority stressors and suicidal ideation and suicide attempts among LGBT adolescents and young adults.

Fox (Vulpes vulpes) involvement identified in a series of cat carcass mutilations.

This study was designed to identify the cause of mutilation and death in 32 cats, part of a larger cohort found dead in Greater London, the United Kingdom, between 2016 and 2018. At the time, discussion in the media led to concerns of a human serial cat killer (dubbed The Croydon Cat Killer) pursuing domestic cats, causing a state of disquietude. Given the link between animal abuse and domestic violence, human intervention had to be ruled out. Using a combination of DNA testing, computed tomography imaging, and postmortem examination, no evidence was found to support any human involvement. Instead, a significant association between cat carcass mutilation and the presence of fox DNA was demonstrated. Gross examination identified shared characteristics including the pattern of mutilation, level of limb or vertebral disarticulation, wet fur, wound edges with shortened fur, and smooth or irregular contours, and marks in the skin, muscle, and bone consistent with damage from carnivore teeth. Together these findings supported the theory that the cause of mutilation was postmortem scavenging by red foxes (Vulpes vulpes). The probable cause of death was established in 26/32 (81%) carcasses: 10 were predated, 8 died from cardiorespiratory failure, 6 from blunt force trauma, one from ethylene glycol toxicity, and another from liver failure. In 6 carcasses a cause of death was not established due to autolysis and/or extensive mutilation. In summary, this study highlights the value of a multidisciplinary approach to fully investigate cases of suspected human-inflicted mutilation of animals.

Mental Health of Lesbian, Bisexual, and Other-Identified Parents and Non-Parents from a Population-Based Study.

Studies have compared sexual minority mothers (mostly lesbian) to heterosexual mothers on mental health, but little research has compared sexual minority women with and without children. This was the first study to compare sexual minority women who did or did not have children, using a population-based sample with three age cohorts. Unlike prior convenience studies, this study finds parents more likely to be bisexual, in a relationship with a man, and non-urban. Bisexual parents scored higher than lesbian parents on psychological distress and lower on life satisfaction and happiness; they also reported less connection to the lesbian, gay, bisexual, and transgender (LGBT) community. Among lesbians, the oldest non-parents reported more happiness and less psychological distress than the youngest non-parents. Parents with other identities perceived more social support from friends and reported lower levels of internalized homophobia than bisexual parents. The results will help professionals and policymakers understand how parenthood status affects women across sexual identities.

Out-of-hours primary care for people with intellectual disabilities: interviews in general practice.

BACKGROUND: Out-of-hours primary care for people with intellectual disabilities (ID) involves different groups of health care professionals, who are often not familiar to one another nor to the specific health care needs of this vulnerable population. It is not known to what extend these specific health care needs or organizational factors influence the delivery of out-of-hours primary care for people with ID. OBJECTIVE: The objective of this study is to explore the experiences of health care professionals regarding the medical content and the organizational context of out-of-hours primary care for people with ID. METHODS: Semi-structured interviews were conducted with daily care professionals, triage nurses and general practitioners (GPs) involved in out-of-hours primary care for people with ID in the Netherlands. Interviews were thematically analysed for medical content and organizational context. RESULTS: The analysis resulted in four interconnected themes: (i) uncertainties in the triage assessment of ID patients; (ii) confusion about inter-professional responsibilities; (iii) impact on routines and workflow concerning ID patients and (iv) constraints in the decision-making process. All issues raised were related to the organizational context. CONCLUSIONS: Health care professionals involved in out-of-hours primary care for people with ID indicate that the quality of this care is more influenced by the organizational context than by the medical content. We recommend out-of-hours GP services and care provider services for people with ID to set standards for roles and responsibilities in order to facilitate health care professionals in delivering accessible and high-quality care to this vulnerable population.

Father-child bonding and mental health in gay fathers using cross-border surrogacy during the COVID-19 pandemic.

RESEARCH QUESTION: What are the psychological implications of the COVID-19 pandemic for father-child bonding and mental health among Italian gay fathers pursuing surrogacy in the USA or Canada? DESIGN: Between 20 March and 29 July 2020, this cross-sectional case-control study collected data on father-child bonding quality, depression, anxiety and somatization in 30 Italian gay fathers (n=15 families) who were having or successfully had a child through cross-border surrogacy during the COVID-19 pandemic. These fathers were compared with a sociodemographically similar group of 50 Italian gay fathers (n=25 families) who had children through cross-border surrogacy prior to the pandemic. RESULTS: Although father-child bonding quality and the mental health symptoms of fathers scored below the clinical cut-off points in both groups, fathers who had or were having a child during the COVID-19 pandemic reported poorer father-child bonding (estimate 3.04, SE 1.47, P=0.044) and more depressive (estimate -1.47, SE 0.49, P=0.005), anxious (estimate -1.96, SE 0.55, P<0.001) and somatic symptoms (estimate -2.48, SE 0.52, P<0.001). CONCLUSIONS: The findings call for the development of international guidelines for cross-border surrogacy and underline the need for tailored and ongoing psychological and legal support for intended gay fathers to ease their strain and anxiety related to having a child through cross-border surrogacy during the COVID-19 pandemic.

Detection of Mycobacterium tuberculosis complex DNA in CD34-positive peripheral blood mononuclear cells of asymptomatic tuberculosis contacts: an observational study.

BACKGROUND: Haematopoietic stem cells expressing the CD34 surface marker have been posited as a niche for Mycobacterium tuberculosis complex bacilli during latent tuberculosis infection. Our aim was to determine whether M tuberculosis complex DNA is detectable in CD34-positive peripheral blood mononuclear cells (PBMCs) isolated from asymptomatic adults living in a setting with a high tuberculosis burden. METHODS: We did a cross-sectional study in Ethiopia between Nov 22, 2017, and Jan 10, 2019. Digital PCR (dPCR) was used to determine whether M tuberculosis complex DNA was detectable in PBMCs isolated from 100 mL blood taken from asymptomatic adults with HIV infection or a history of recent household or occupational exposure to an index case of human or bovine tuberculosis. Participants were recruited from HIV clinics, tuberculosis clinics, and cattle farms in and around Addis Ababa. A nested prospective study was done in a subset of HIV-infected individuals to evaluate whether administration of isoniazid preventive therapy was effective in clearing M tuberculosis complex DNA from PBMCs. Follow-up was done between July 20, 2018, and Feb 13, 2019. QuantiFERON-TB Gold assays were also done on all baseline and follow-up samples. FINDINGS: Valid dPCR data (ie, droplet counts >10 000 per well) were available for paired CD34-positive and CD34-negative PBMC fractions from 197 (70%) of 284 participants who contributed data to cross-sectional analyses. M tuberculosis complex DNA was detected in PBMCs of 156 of 197 participants with valid dPCR data (79%, 95% CI 74-85). It was more commonly present in CD34-positive than in CD34-negative fractions (154 [73%] of 197 vs 46 [23%] of 197; p<0·0001). Prevalence of dPCR-detected M tuberculosis complex DNA did not differ between QuantiFERON-negative and QuantiFERON-positive participants (77 [78%] of 99 vs 79 [81%] of 98; p=0·73), but it was higher in HIV-infected than in HIV-uninfected participants (67 [89%] of 75 vs 89 [73%] of 122, p=0·0065). By contrast, the proportion of QuantiFERON-positive participants was lower in HIV-infected than in HIV-uninfected participants (25 [33%] of 75 vs 73 [60%] of 122; p<0·0001). Administration of isoniazid preventive therapy reduced the prevalence of dPCR-detected M tuberculosis complex DNA from 41 (95%) of 43 HIV-infected individuals at baseline to 23 (53%) of 43 after treatment (p<0·0001), but it did not affect the prevalence of QuantiFERON positivity (17 [40%] of 43 at baseline vs 13 [30%] of 43 after treatment; p=0·13). INTERPRETATION: We report a novel molecular microbiological biomarker of latent tuberculosis infection with properties that are distinct from those of a commercial interferon-γ release assay. Our findings implicate the bone marrow as a niche for M tuberculosis in latently infected individuals. Detection of M tuberculosis complex DNA in PBMCs has potential applications in the diagnosis of latent tuberculosis infection, in monitoring response to preventive therapy, and as an outcome measure in clinical trials of interventions to prevent or treat latent tuberculosis infection. FUNDING: UK Medical Research Council.

Epidemiology of Bovine Tuberculosis and Its Zoonotic Implication in Addis Ababa Milkshed, Central Ethiopia.

Bovine tuberculosis (bTB) continues to be one of the most widely distributed chronic infectious diseases of zoonotic importance, which causes a significant economic loss in animal production. A cross-sectional study was conducted to estimate the prevalence of bTB and its associated risk factors and type the Mycobacterium bovis isolated in central Ethiopia. A total of 65 dairy farms and 654 cattle were tested for bTB using a single intradermal comparative cervical tuberculin (SICCT) test. Data on farm management, animal-related characteristics, and the owner's knowledge of the zoonotic importance of bTB were collected using a structured questionnaire. In addition, a total of 16 animals from different farms were identified for postmortem examination. Lowenstein Jensen (LJ) culture was also conducted, and spoligotyping was used to type the M. bovis strains isolated. Chi-square test and logistic regression models were used to analyze the herd- and animal-level risk factors. Herd- and animal-level prevalence rates of bTB were 58.5% (95% CI: 46.2%-69.2%) and 39.3% (95% CI: 35.5%-43.5%), respectively. At the herd level, poor farm management was the predictor for bTB positivity (p < 0.05). Animal breed, poor BCS, farm type, and poor farm management conditions were significant predictors of bTB positivity (p < 0.05) at an individual animal level. All animals identified for postmortem examination were found to have gross TB-like lesions. A total of 14 M. bovis strains were identified from 12 animals that were positive for LJ culture. The strain with the largest number of clusters (five isolates) was SB1176, followed by SB0134 (three isolates), SB0192 (two isolates), and SB2233 (two isolates), and two new strains, each consisting of only one isolate. The majority (58.5%) of the respondents did not know the zoonotic importance of bTB. The result of this study showed a high prevalence of bTB in the Addis Ababa milkshed and a low level of consciousness of the owners on its transmission to humans. Therefore, the launching of acceptable control measures of bTB and the creation of public awareness about its zoonotic transmission and prevention measures are required.