OBJECTIVE: The queen is recognised as an induced ovulator. Ovulation without male contact is generally regarded as spontaneous. The aim of this study was to provide an estimate of the incidence of spontaneous ovulation in a population of intact queens presented to a veterinary care facility for both reproductive and non-reproductive reasons. The secondary objective was to determine the roles of age, breed, body weight, presence of tom cats or other cycling queens, and physical contact with humans on triggering spontaneous ovulation, along with its implications.
METHODS: Serum samples from post-pubertal intact queens presented between January 2020 and June 2023 to the Veterinary Teaching Hospital of the University of Padova, Italy, were retrieved and assayed for progesterone (P4) levels. Serum P4 above 2.0 ng/ml without a history of male contact was considered as proof of spontaneous ovulation.
RESULTS: In total, 31 serum samples from 29 intact post-pubertal queens were obtained. Of the 31 samples, 14 had a P4 concentration above 2.0 ng/ml and 9/29 (31.0%) queens ovulated spontaneously. The mean age and weight of the nine spontaneously ovulating queens were 4.3 ± 5.7 years and 3.7 ± 0.8 kg, respectively. One queen ovulated spontaneously at her first heat at 6 months of age, which makes it the earliest spontaneous ovulation reported.
CONCLUSIONS: As both our findings and previous publications indicate that the incidence of spontaneous ovulation in queens is consistently ⩾30%, cats should not be considered strictly induced ovulators, but as a species in which ovulation can be either spontaneous or induced. Since the risk of progesterone-dependent conditions (cystic endometrial hyperplasia - pyometra complex, feline mammary hypertrophy) is increased in these queens, veterinarians should be aware and advise breeders and clients accordingly.
Female cats ovulate upon vaginal stimulation exerted by the spikes of the male’s penis while mating, which makes them induced ovulators. When ovulation occurs without male contact, it is considered spontaneous. There are several factors that are thought to facilitate this non-induced ovulation, but no consensus on their relevance. The aim of this study was to provide an estimate of the rate of spontaneous ovulation in a population of intact female cats of various breeds presented to a veterinary care facility, as well as the influence of factors such as age, breed, body weight, presence of male cats or other cycling females, and physical contact with humans on triggering spontaneous ovulation. In addition, possible implications arising from progesterone exposure were assessed.Progesterone was retrospectively assayed in the serum of adult cycling female cats presented to the Veterinary Teaching Hospital of the University of Padova, Italy, between January 2020 and June 2023. Values above 2.0 ng/ml without a history of male contact were considered proof of spontaneous ovulation. Out of 29 cats, nine (31%) ovulated spontaneously, with one female having done so at puberty (6 months of age), which makes it the first spontaneous ovulation ever reported in a pubertal queen.As spontaneous ovulation has been found to occur at a rate of more than 30% both in our and in previous publications on this topic, we propose that cats should be considered both an induced and a spontaneously ovulating species. Since animals that ovulate spontaneously, and therefore experience additional luteal phases, are at a higher risk of developing progesterone-dependent conditions, veterinarians should be aware and advise breeders and clients accordingly.

AbstractThe common eastern bumble bee (Bombus impatiens) queens endure cold winter months by entering a diapause state. During this overwintering period, these animals use stored energy reserves while maintaining a low metabolic rate. This study investigates changes in the metabolic rate of bumble bee queens during diapause-like laboratory conditions and the potential reorganization of the flight muscle metabolic properties during this period. We first confirmed the hypometabolic state of queens during diapause in the laboratory, which lowered their resting metabolic rate to less than 5% of normal resting values. Body mass decreased during diapause, body composition changed where carbohydrates decreased initially, and later protein declined, with a similar trend for lipid content. Using cellular respirometry, we determined the capacity of the flight muscle cells of bumble bee queens to use various metabolic fuels and whether this capacity changes during the progression of diapause to favor stored lipid-derived substrates. Queens showed a low capacity to oxidize the amino acid proline, compared with workers, and their capacity to oxidize all metabolic substrates did not change during a 4-mo diapause period in the laboratory. We also show no detectable ability to oxidize fatty acid by flight muscle mitochondria in this species. The metabolic properties of flight muscle tissue were further characterized using metabolic enzyme activity profiles showing little change during diapause, indicating that profound metabolic suppression is induced without major changes in muscle metabolic phenotypes. Overall, B. impatiens queens undergo diapause while maintaining flight muscle capacity under the conditions used.

The health of honey bee queens is crucial for colony success, particularly during stressful periods like overwintering. To accompany a previous longitudinal study of colony and worker health, we explored niche-specific gut microbiota, host gene expression, and pathogen prevalence in honey bee queens overwintering in a warm southern climate. We found differential gene expression and bacterial abundance with respect to various pathogens throughout the season. Biologically older queens had larger microbiotas, particularly enriched in Bombella and Bifidobacterium. Both Deformed Wing Virus A and B subtypes were highest in the fat body tissue in January, correlating with colony Varroa levels, and Deformed Wing Virus titers in workers. High viral titers in queens were associated with decreased vitellogenin expression, suggesting a potential trade-off between immune function and reproductive capacity. Additionally, we found a complex and dynamic relationship between these viral loads and immune gene expression, indicating a possible breakdown in the coordinated immune response as the season progressed. Our study also revealed a potential link between Nosema and Melissococcus plutonius infections in queens, demonstrating that seasonal opportunism is not confined to just workers. Overall, our findings highlight the intricate interplay between pathogens, metabolic state, and immune response in honey bee queens. Combined with worker and colony-level metrics from the same colonies, our findings illustrate the social aspect of queen health and resilience over the winter dearth.

Feline breeding colonies are important to the feline industry by preserving traits desirable for a particular breed or in research settings by maintaining medically valuable genetic traits. As breeding females age, their reproductive efficiency declines. The objective of this study was to determine the most common causes of infertility in breeding females that were producing fewer kittens. Knowing the cause and average age of infertility would allow management decisions to be made for the betterment of the colony. The medical records of 70 queens retired from breeding from a single research colony were examined for litter size and number, fertility over their lifespan, and age and reason for removal from breeding stock. Sections of uterus and ovaries were evaluated using gross and histopathological examination for a subset of these queens (46). The data suggests that mature, continuously breeding female cats may show signs of reduced fertility (infertile matings or reduced litter size) as early as 3 years of age and may be a result of undiagnosed Cystic Endometrial Hyperplasia (CEH), endometritis, pyometra and/or ovarian cysts. Evaluation of breeding queens should include periodic ultrasounds to monitor for ovarian cysts and evidence of CEH. Retiring animals from breeding once signs of infertility become apparent is recommended.

Host symbionts are often considered an essential part of the host phenotype, influencing host growth and development. Bumble bee is an ideal model for investigating the relationship between microbiota and phenotypes. Variations in life history across bumble bees may influence the community composition of gut microbiota, which in turn influences phenotypes. In this study, we explored gut microbiota from four development stages (early-instar larvae, 1st instar; mid-instar larvae, 5th instar; late-instar larvae, 9th instar; and adults) of workers and queens in the bumble bee Bombus terrestris using the full-length 16S rRNA sequencing technology. The results showed that morphological indices (weight and head capsule) were significantly different between workers and queens from 5th instar larvae (p < 0.01). The alpha and beta diversities of gut microbiota were similar between workers and queens in two groups: early instar and mid instar larvae. However, the alpha diversity was significantly different in late instar larvae or adults. The relative abundance of three main phyla of bacteria (Cyanobacteria, Proteobacteria, and Firmicutes) and two genera (Snodgrassella and Lactobacillus) were significantly different (p < 0.01) between workers and queens in late instar larvae or adults. Also, we found that age significantly affected the microbial alpha diversity as the Shannon and ASVs indices differed significantly among the four development stages. Our study suggests that the 5th instar larval stage can be used to judge the morphology of workers or queens in bumble bees. The key microbes differing in phenotypes may be involved in regulating phenotypic variations.

Pollinators have to cope with a wide range of stressful, not necessarily lethal factors limiting their performance and the ecological services they provide. Among these stressors are pesticides, chemicals that are originally designed to target crop-harming organisms but that also disrupt various functions in pollinators, including flight, communication, orientation and memory. Although all these functions are crucial for reproductive individuals when searching for mates or nesting places, it remains poorly understood how pesticides affect reproduction in pollinators. In this study, we investigated how a widely used fungicide, boscalid, affects reproduction in honey bees (Apis mellifera), an eusocial insect in which a single individual, the queen, fulfills the reproductive functions of the whole colony. Boscalid is a succinate dehydrogenase inhibitor (SDHI) fungicide mainly used on rapeseed flowers to target mitochondrial respiration in fungi but it is also suspected to disrupt foraging-linked functions in bees. We found that immature queen exposure to sublethal, field relevant doses of boscalid disrupted reproduction, as indicated by a dramatic increase in queen mortality during and shortly after the nuptial flights period and a decreased number of spermatozoa stored in the spermatheca of surviving queens. However, we did not observe a decreased paternity frequency in exposed queens that successfully established a colony. Queen exposure to boscalid had detrimental consequences on the colonies they later established regarding brood production, Varroa destructor infection and pollen storage but not nectar storage and population size. These perturbations at the colony-level correspond to nutritional stress conditions, and may have resulted from queen reduced energy provisioning to the eggs. Accordingly, we found that exposed queens had decreased gene expression levels of vitellogenin, a protein involved in egg-yolk formation. Overall, our results indicate that boscalid decreases honey bee queen reproductive quality, thus supporting the need to include reproduction in the traits measured during pesticide risk assessment procedures.

Domestic felids (Felis catus) have been traditionally categorized as seasonal polyestrous with induced ovulation. Thus, the ability to augment or distribute the number of litters born throughout the year would offer a desirable advantage. Artificial-light regimens have been used to overcome seasonal variations in this species. Understanding the mechanisms that underlie photoperiodicity might enable the development of improved and sustainable breeding schemes. The aim of this article was therefore to summarize the present knowledge on the effect of artificial light on female-cat reproduction. To that end a systematic review of the literature from 1940 to the present was performed. International original articles and scientific abstracts were also included, and at the conclusion we emphasized areas that require further research.

Honey bee queens show extreme fecundity, commonly laying more than a thousand eggs in a single day. It has proven challenging to study the temporal organization of egg-laying behavior because queens are typically active around the clock in the dark cavity of a densely populated nest. To contend with this challenge, we developed two novel methods allowing detailed monitoring of queen activity and egg laying. We first adapted a high-resolution, continuous, tracking system allowing to track the position of barcode-tagged queens in observation hives with colonies foraging outside. We found that the queen is active ~96% of the day with typically no diurnal rhythm. Next, we developed a new laboratory procedure to monitor egg laying at single egg resolution under different light regimes. We found that under constant darkness (DD) and temperature conditions, queens laid eggs with no circadian rhythms. Queen fecundity was severely reduced under constant light (LL). Under a 12:12 illumination regime, queen fecundity was comparable to under constant darkness, with a higher number of eggs during the light phase. These daily rhythms in egg laying continued when these queens were released to DD conditions, suggesting that egg-laying rhythms are influenced by endogenous circadian clocks. These results suggest that honey bee queens are active and lay eggs around the clock with no diurnal rhythms. Light has complex influences on these behaviors, but more studies are needed to determine whether these effects reflect the influence of light directly on the queen or indirectly by affecting workers that interact with the queen.

Social insect societies are characterized by a high level of organization. This is made possible through a remarkably complex array of pheromonal signals produced by all members of the colony. The queen\'s pheromones signal the presence of a fertile female and induce daughter workers to remain sterile. However, the lack of the queen mandibular pheromone leads to the emergence of rebels, i.e., workers with increased reproductive potential. We suggested that the rebels would have developed tergal glands and reduced Nasonov glands, much like the queen but contrary to normal workers. Our guess turned out to be correct and may suggest that the rebels are more queen-like than previously thought. The tergal gland cells found in the rebels were numerous but they did not adhere as closely to one another as they did in queens. In the rebels, the number of Nasonov gland cells was very limited (from 38 to 53) and there were fat body trophocytes between the glandular cells. The diameters of the Nasonov gland cell nuclei were smaller in the rebels than in the normal workers. These results are important for understanding the formation of the different castes of Apis mellifera females, as well as the division of labor in social insect societies.

The proteolytic and antioxidant systems are important components of humoral immunity, and these biomarkers indicate the immune status. These compounds are synthesized in the bees\' fat body and released into the hemolymph. Their functions maintain the organism\'s homeostasis and protect it against adverse environmental factors (including pathogens). We determined the activities of acidic, neutral, and alkaline proteases and their inhibitors, as well as superoxide dismutase (SOD), catalase (CAT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and the level of total antioxidant potential (TAC). These compounds were investigated in the fat body and hemolymph in the females with increased reproductive potential, i.e., queens and rebels, and in normal (non-reproductive sterile non-rebel) workers. The phenoloxidase (PO) activities were determined in the hemolymph. The normal workers had higher activities of proteases and their inhibitors, SOD and CAT, in the fat body and hemolymph, compared to the queens and rebels. The protease inhibitors were not usually active in the queens. As we predicted, the rebels revealed values between those of the queens and normal workers. The highest activities of proteases and antioxidants were identified in the fat body from the third tergite in comparison with the sternite and the fifth tergite. These results are important for oxidative stress ecology and give a better understanding of the functioning of the fat body and the division of labor in social insects.