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Animals

MDPI AG

All preprints, ranked by how well they match Animals's content profile, based on 23 papers previously published here. The average preprint has a 0.03% match score for this journal, so anything above that is already an above-average fit. Older preprints may already have been published elsewhere.

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SUCCESSFUL IN VITRO EMBRYO PRODUCTION WITH OOCYTES ASPIRATED FROM LIVE WHITE-TAILED DEER (Odocoileus virginianus texanus) DONNORS UNDER CAPTIVITY IN NORTHEAST MEXICO

MarabotoM., E. E.; Trejo M., F. J.; Del Angel R., H.; Rosales H., J.; Bautista M., Y.; Torres P., L. I.; Gonzalez-Reyna, A.

2022-02-25 zoology 10.1101/2022.02.18.481106 medRxiv
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Assisted reproductive technologies (ARTs), such as artificial insemination, semen sorting and freezing, embryo production in vivo and in vitro, are all methods in which animal reproduction management of several species has become considerably more efficient. These ARTs have been applied both in domestic and wild ruminants. In this study, in vitro embryo production was attempted with oocytes collected surgically during the mating season, from live white-tailed deer (WTD) hinds, maintained under captivity, in Northeast Mexico. The study was conducted in two WTD farms nearby Cd. Victoria, Tamps., Mexico, and another WTD farm located in Guerrero Coahuila, Mexico. The laboratory work was carried at the Centro de Desarrollo de la Capacidad Productiva y Mejoramiento Genetico de la Ganaderia, property of the Union Ganadera Regional de Tamaulipas (Centro-UGRT). The deer hinds are kept in captivity year-around, with feeding and health management provided accordingly to their requirements (50-60 kg females). Fresh green forage and clean fresh water is supplied daily. Oocytes were collected on each farm, while the hinds were maintained under general anesthesia and by means of a mid-ventral laparotomy, ovaries were exposed and follicles greater than 1-2 mm were aspirated with a 20 G needle, connected by a two-way plastic and latex hose to a vacum machine (WTA, Brazil); the oocytes were collected into a 50 ml. centrifuge tube containing wash media with heparine (Vitrogen, Brazil). Each oocyte collection lasted approximately 20 minutes, after which and while still on the farm, the oocytes were filtered with 50 {micro} mesh filters and rinsed several times with wash media, then placed on search Petri plates. Oocytes were then counted and classified (total, viable and non-viable oocytes) and placed into cryovials with maturation media (3 ml. MIV, Vitrogen, Brazil) and placed into a portable incubator with 5 % CO2 gas mix (LabiMixWTA, Brazil), after all hinds were done, oocytes were transported to the in vitro fertilization laboratory (IVF Lab) at the Centro-UGRT. Once at the IVF Lab, the oocytes were placed on a larger incubator (Eve, WTA, Brazil) and kept there for 18-20 hours, after which, the MIV media was changed for fertilization media (FIV, Vitrogen, Brazil) and fertilization was initiated by adding 10,000-12,000 live sperm per cryovial, and incubated for an additional period of 24 hours; after which, FIV media was replaced by the same media and at this point, cleavage rate was estimated by counting the oocytes that initiated cell division. At 72 hours after cell division started, fertilization rate was estimated; and 7 days after, the blastocysts were counted and classified. The whole process of oocyte maturation and embryo production in vitro, was conducted based on a beef cattle embryo production system and adapted to a deer embryo production system using media for small ruminants (Vitrogen, Brazil). Data collected per hind included total number of oocytes, viable and non-viable oocytes, cleavage rate (ratio of viable oocytes that initiated cell division over viable oocytes), fertilization rate (ratio of embryos that initiated cleavage over those that continued development to the blastocyst stage) and blastocyst rate (embryos reaching the blastocyst stage over cleaved embryos); averages were calculted for each parameter. The main results from this study on a per hind basis for total viable and non-viable oocytes were 9.8, 6.3 and 4.5, respectively; cleavage and blastocyst rates were 39.5 and 36.8, respectively and 2.3 blastocysts. In conclusion, oocyte collection from live WTD hinds and in vitro embryo production was succesfully done under farming conditions in Northeast Mexico.

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Pathologies and causes of death in stranded cetaceans in the Canary Islands (2013-2018)

Diaz Santana, P. J.; Arbelo, M.; Diaz-Delgado, J.; Groch, K.; Suarez-Santana, C.; Consoli, F.; Bernaldo de Quiros, Y.; Quesada-Canales, O.; Sierra, E.; Fernandez, A.

2026-04-05 pathology 10.64898/2026.04.01.715953 medRxiv
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Cetacean pathology is a cornerstone for population and marine ecosystem health monitoring, allowing clear differentiation among natural and anthropogenic threats. Previous studies in the Canary Islands reported natural causes of death in 59.4% (1999-2005) and 81% (2006-2012) of stranded cetaceans, versus anthropogenic causes in 33.3% and 19%, respectively. This study aimed to determine the causes of death (CD), pathologic findings, and epidemiological patterns of 316 cetaceans stranded in the Canary Islands between 2013 and 2018. The CDs were classified in pathologic entities (PEs) emphasizing natural versus anthropic origins. Of 316 animals, 224 (70.9%) from 18 species were suitable for pathological investigations. Among natural PEE, natural pathology associated with good nutritional status (NP-GNS) and natural pathology associated with significant loss of nutritional status (NP-LNS) represented 43/224 (19.2%) and 36/224 (16%) cases, respectively. Natural pathology with undetermined nutritional status (NP-UNS) occurred in 19/224 (8.5%) animals. Intra- and interspecific traumatic interactions (ITI) represented 30/224 (13.4%) cases, followed by neonatal/perinatal pathology (NPN) 19/224 (8.5%) and live-stranding stress and/or capture myopathy (LS-CM) 18/224 (8%). Infectious and parasitic diseases predominated in natural PEs. Anthropogenic PEs included interaction with fishing activities (IFA) in 17/224 (7.6%) cases, vessel collisions (VC) in 9/22 (4%) cases, and foreign body-associated pathology (FBAP) in 3/224 (1.3%) animals. Overall, anthropogenic causes accounted for 12.9% of deaths, natural causes for 73.6%, and the CD could not be established in 30/194 (13.4%) cases. This study reaffirms the trends concerning recognized PEs (NP-GNS, NP-LNS, NP-UNS, ITI, NPN, LS-CM, IFA, VC, and FBAP), expands the body of knowledge on cetacean pathology in the Canary Islands, and reports novel findings including mixed infections, clostridiosis in uncommon species, uremic syndrome secondary to urethral nematodiasis, gas embolism in unusual species, epibiont stomatitis, congenital musculo-skeletal malformations, or neoplastic processes. These findings advance understanding of cetacean mortality patterns and support conservation and management strategies.

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Effects of periodic group housing opportunities on reproductive performance and welfare in sows

Shimasaki, T.; Yoyou, K.; Kojima, T.; Huang, C.-Y.; Kato, H.; Ishida, M.; Takeda, K.

2026-05-21 animal behavior and cognition 10.64898/2026.05.19.726187 medRxiv
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ObjectiveStall housing of pregnant sows raises welfare concerns, whereas conventional group housing systems often reduce space efficiency. This study evaluated the effects of periodic group housing (PG) on reproductive performance and welfare compared with continuous stall housing (CS). MethodsSows in the CS group (n = 15) were continuously housed in stalls. In the PG group (n = 15), sows were housed in groups of three and allocated 1 day of group housing and 6 days of stall housing per week over 10 weeks. During group housing sessions, the sows had access to a group housing area containing sawdust. Behavioral observations and salivary cortisol measurements were conducted on the first day of the stall housing session in weeks 1, 6, and 10. Behavioral indices were expressed as proportions based on 90 sampling points recorded at 1-min intervals. ResultsThe number of stillbirths was significantly lower in the PG group than in the CS group (0.63 vs. 1.49 piglets per litter). whereas other reproductive outcomes, including total litter size and average birth weight, did not differ. In older parity sows, the PG treatment markedly increased the proportion of time spent lying, suggesting reduced discomfort associated with restricted movement. Furthermore, the proportion of exploratory behavior decreased markedly, and drinking behavior showed a decreasing trend across parity levels in the PG group, suggesting partial satisfaction of motivations for environmental exploration and oral manipulation. The proportion of oral abnormal behavior showed a pronounced interaction between housing treatment and experimental week, increasing from week 1 to week 6 in the PG group. Salivary cortisol concentrations did not differ between the groups. ConclusionPG may improve reproductive performance and partially satisfy the behavioral motivations restricted under continuous stall housing. This system may represent a practical alternative for improving animal welfare while minimizing economic losses.

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In vitro fertilisation procedure assisted with computer vision models for organic Senegalese sole (Solea senegalensis) culture

Qadir, A.; Martinez, S. S.; Serratosa, F.; Duncan, N.

2026-03-02 animal behavior and cognition 10.64898/2026.02.26.707220 medRxiv
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Reproductive dysfunction remains a major challenge for Senegalese sole (Solea senegalensis) aquaculture. Hormone-induced ovulation and in vitro fertilisation (IVF) are currently used to overcome the absence of natural courtship behaviours in captivity. This study investigates the feasibility of hormone-free IVF and behaviour-based prediction of ovulation as alternative strategies to enhance reproductive outcomes. We selected males using computer-assisted sperm analysis to assess sperm motility and quality for IVF trials. IVF trials were conducted using selected males and naturally ovulated eggs collected from females during evening hours across six experimental nights in two groups. Fish behaviour was continuously recorded using underwater cameras, and a convolutional neural network was developed to automatically detect Rest the Head (RTH) and Locomotor Activity (LA) behaviours. These behavioural counts, together with timing information, were used as features to train a logistic regression model for predicting ovulation events. Hormone-free IVF achieved fertilization rates up to 44% with 18% hatching success, producing viable larvae without hormonal intervention. Both groups showed significantly elevated RTH and LA during ovulation nights compared to non-ovulation nights, with peak activity occurring between 18:00-19:00 hours. The behavioural prediction model correctly identified ovulation with 82-85% accuracy and an area under the curve of 0.95. These findings demonstrate that sperm-quality-based male selection combined with automated behaviour analysis provides a practical, non-invasive approach for hormone-free reproduction in organic flatfish aquaculture.

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Obesity status and its relative factors of captive Asian elephants (Elephas maximus) in China based on body condition assessment

Tang, Y.; Jia, T.; Zhou, F.; Wang, L.; Zhang, L.

2023-11-25 zoology 10.1101/2023.11.25.568637 medRxiv
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Obesity is a common health problem in captive wildlife. Since the obesity status of captive Asian elephants (Elephas maximus) in China has not previously been investigated, we recorded seven relevant variables (sex, age, daily feed supply, proportion of high-calorie feed, outdoor enclosure area, outdoor time, and foot disorder) related to obesity in 204 captive Asian elephants through field investigation of 43 elephant-raising facilities. Assessment of obesity was based on visual body condition scoring for each individual. It revealed that obesity was prevalent for captive Asian elephants (especially elephants in zoos) in China. Over 70% of captive Asian elephants in China were overweight or in obesity to various degrees. Statistical analysis showed that for elephants in zoos, insufficient outdoor time might be the primary potential cause of obesity. We suggested facilities to extend the outdoor time and control the supply of high-calorie feed (e.g., fruits, vegetables, pellets, etc.) for captive elephants, thereby alleviating obesity through increased exercise and a suitable energy intake. Moreover, all facilities should implement positive reinforcement training to facilitate regular physical examinations, including foot health checks and blood sampling. This training would improve the ability to collect more precise information relating to elephant health, obesity, and the evaluation of animal welfare.

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Dairy vs. beef production - expert views on welfare

Mandel, R.; Bracke, M. B. M.; Nicol, C. J.; Webster, J.; Gygax, L.

2022-01-20 animal behavior and cognition 10.1101/2021.12.06.471462 medRxiv
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Consumers views and concerns about the welfare of farm animals may play an important role in their decision to consume dairy, meat and/or plants as their primary protein source. As animals are killed prematurely in both dairy and beef industries, it is important to quantify and compare welfare compromise in these two sectors before the point of death. Seventy world-leading bovine welfare experts based in 23 countries, were asked to evaluate the likelihood of a bovine to experience 12 states of potential welfare concern, inspired by the Welfare Quality(R) protocol. The evaluation focused on the most common beef and dairy production systems in the experts country, and was carried out separately for dairy/beef calves raised for red-meat, dairy/beef calves raised for veal, dairy/beef calves raised as replacement, and for dairy/beef cows. The results show experts rated the overall likelihood of a negative welfare state (i.e. welfare risk) to be higher in animals from dairy herds than from beef herds, for all animal categories, regardless of whether they were used to produce milk, red-meat or veal. These findings suggest that consuming food products derived from common dairy production systems (dairy or meat), may be more harmful to the welfare of animals than consuming products derived from common beef production systems (i.e. from animals solely raised for their meat). Raising awareness about the linkage between dairy and meat production, and the toll of milk production on the welfare state of animals in the dairy industry, may encourage a more sustainable and responsible food consumption.

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Ocular Disease Prevalence in California Sea Lions In Human Care: Freshwater Versus Saltwater Housing

Brehm, I.; Herzner, S.; Baumgartner, K.; Beckmann, J.; Simon, R.; von Fersen, L.

2024-12-22 zoology 10.1101/2024.12.20.629697 medRxiv
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California sea lions (Zalophus californianus), are susceptible to various ocular diseases, with some literature suggesting a heightened risk for those kept in freshwater pools as opposed to saltwater. Given the potential implications of housing conditions on animal health, we wanted to analyze the ocular health of two distinct groups of sea lions kept at Nuremberg Zoo, one housed in freshwater and the other in saltwater. Data extracted from the animals medical records over a ten-year period were used to compare the incidence of eye conditions and other medical conditions observed. The results revealed no significant difference in the overall prevalence of eye diseases between the two environments. However, a distinct seasonal pattern was noted: sea lions kept in freshwater exhibited a peak in eye disorders during the summer months, while those in saltwater displayed a more uniform distribution of the occurrence of ocular diseases throughout the year. These findings suggest that sun exposure and water quality are potentially more influential factors in the development of ocular diseases in pinnipeds than salinity or UV radiation. Further studies are necessary to elucidate the underlying mechanisms and optimize care practices for these marine mammals. Simple SummaryThis study investigated the ocular health of California sea lions (Zalophus californianus) at Nuremberg Zoo, comparing those housed in freshwater pools to those in saltwater. An analysis of medical records over ten years showed no significant difference in the overall prevalence of eye diseases between the two groups. However, sea lions in freshwater experienced a peak of eye disorders in the summer, while those in saltwater had a more consistent occurrence throughout the year. These results imply that factors such as sun exposure and water quality may play a more crucial in ocular disease development than salinity or UV radiation. The study highlights the need for further research to understand these dynamics and improve care for marine mammals.

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Comparison of different head and neck positions and behaviour in ridden elite dressage horses between warm-up and competition

Kienapfel, K.; Bachmann, I.; Piccolo, L.; Ruess, D.; Gmel, A.

2021-12-17 animal behavior and cognition 10.1101/2021.12.17.473217 medRxiv
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Head and neck position (HNP) has been identified in literature as important influence on wellbeing. It was investigated in ridden elite dressage horses whether there is a relation between the HNP, ethological indicators and the grading in the warm-up area and in the test. 49 starters (83%) of a Grand-Prix Special (CDIO5 *) as part of the CHIO in Aachen 2018 and 2019 were examined. For each horse-rider pair, HNP (angle at vertical (AT), poll angle (PA), neck angel (NA)) used were analysed as well as conflict behavior for 3 minutes each in warm-up area and test. 6571 individual frames were used. The noseline was carried significantly less behind the vertical in test vs. warm-up (5.43 {degrees} {+/-} 4.19 vs. 11.01 {degrees} {+/-} 4.54 behind the vertical; T = 34.0; p < 0.05). The horses showed significantly less conflict behavior in the test vs. warm-up (123 {+/-} 54 vs. 160 {+/-} 75) (T = 76.00; p < 0.01). In the latter, a smaller PA and more defensive behaviour of the horses was observed compared to the test. A correlation between the grading of test and HNP was found (R = 0.38; p < 0.05). The further the noseline was behind the vertical, the higher was the chance of a good rating. The higher riders were ranked in the "FEI world ranking", the higher were their marks in the competition (2018: r = -0.69, p < 0.05; 2019: r = -0.76, p < 0.05). Horses of riders higher in world ranking tended to show more unusual oral behaviour (r = -0.30, p < 0.05), and a noseline stronger behind the vertical (r = - 0.37, p < 0.05) resulting in a smaller NA (r = 0.43, p < 0.05). This are from the point of view of animal welfare problematic results.

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Morphometric Characterization and Variation of Somali Dromedary Camels (Camelus dromedarius) in a selected district of Benadir Region

Barre, A.

2026-04-15 animal behavior and cognition 10.64898/2026.04.12.718019 medRxiv
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Camels are one of the hardy animal species known for their high adaptability to extreme climatic conditions. However, scientific studies on this important animal species are limited. This study aims to investigate the statistical relationships between some body measurements of dromedary camels with different characteristics raised in the Mogadishu, Somalia. The research was carried out on a total of 248 camels (102 males, 146 females) of different age groups, belonging to the SiifDacar, Hoor, and Eyddimo types, raised in camel farms in Dharkenley, Dayniile, and Hodan districts of Mogadishu. Body measurements were taken manually using a standard measuring tape, and the collected data were subjected to statistical analysis. As a result of the analyses, significant differences were found between the camels named with the local language as mentioned above in terms of withers height, rump height, body length, and shoulder width in female camels. In male camels, it was determined that there was a statistically significant difference among the camels named in terms of rump width. When evaluated specifically for female camels, it was found that the camels of the SiifDacar had higher withers and rump height compared to the camels of the Hoor, and were also longer than the other two different camels. Although no notable difference in shoulder width was found between the Hoor and Eyddimo camels, Siif Dacar females exhibited broader shoulders than both. In male camels, Siif Dacar had a significantly wider rump compared to those of the Hoor camels. Furthermore, the study identified varying degrees of statistically significant correlations among different body measurements in the camels. Overall, it was observed that female camels of the Siif Dacar were larger in size compared to those of the Hoor and Eyddimo camels, and that male camels generally exhibited greater body dimensions than females. These findings contribute to the registration of the different-named camel types in Somalia as separate breeds and to a better understanding of the morphological characteristics of camels.

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A free classified advertising website is a high-risk unregulated source of domestic wild boar, pot-bellied pigs, and their hybrids with domestic swine in Canada: Implications for African Swine Fever Risk Preparedness

MacDonald, A. M.; Brook, R. K.

2022-03-16 scientific communication and education 10.1101/2022.03.14.483774 medRxiv
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Invasive wild pigs are a global problem and are established in large areas across Canada. Domestic European wild boar were first introduced to Canadian farms in the 1980s, and their escape and intentional release have since led to free-ranging populations, which have expanded rapidly in numbers and range. Wild pigs are associated with widespread damage to crops and natural environments, as well as risks of disease transmission. African Swine Fever (ASF) is a highly contagious viral disease of global concern, that can spread between domestic and free-ranging swine and cause catastrophic economic impacts. As ASF preparedness requires a detailed understanding of the distribution and movements of all pig types, this paper aimed to characterize an unregulated grey swine market in Canada to understand risks to domestic swine production and potential contributions to free-ranging wild pig populations. Kijiji.ca is a free Canadian internet classified advertising service, almost exclusively cash-based with no receipts and few, if any, records kept of transactions. We monitored Kijiji for sales of domestic wild boar, pot-bellied pigs, and their hybrids across Canada over two months from April 28 - June 30, 2021. Data was collected from all advertisements, including how the seller labelled pigs breed, age, sex, number for sale, sexual intactness, presence of tattoos and/or ear tags, as well as the date and location of the posting. Locations were mapped and compared to the spatial distribution of existing free-ranging wild pigs in Canada to identify areas they were likely to facilitate new populations or potentially supplement and genetically diversify existing populations. We identified 151 advertisements on Kijiji, the most coming from Ontario (34.4%; n = 52) and Alberta (29.1%; n = 44), followed by Saskatchewan (11.3%; n = 17), British Columbia (10.6%; n = 16), Nova Scotia (7.3%; n = 11), New Brunswick (4.0%; n = 6), Manitoba (2.0%; n = 3), Quebec (0.7%; n = 1), and Prince Edward Island (0.7%; n = 1). No listings were observed in Newfoundland and Labrador, or Yukon, Nunavut and Northwest Territories. Overall, 62 (41%) of advertisements were within watersheds with existing wild pigs that may contribute to supplementing additional swine, and 89 (59%) were within watersheds where wild pigs have not yet been identified. African swine fever preparedness should include policies and action to address this unmonitored and unregulated sale of swine in Canada, including ear tags, tattoos, genetic analysis, and mandatory reporting.

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Vibrio cholerae may be transmitted to humans from bullfrog through food or water

yang, y.; zhu, x.; chen, y.; liu, y.; song, y.; ai, x.

2021-04-09 pathology 10.1101/2021.04.09.439145 medRxiv
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Bullfrog is one of the most important economic aquatic animals in China. It is widely cultured in southern China, and is a key breed recommended as an industry of poverty alleviation in China. During recent years, a fatal bacterial disease has often been found in cultured bullfrogs. The clinical manifestations of the diseased bullfrogs were severe intestinal inflammation and even anal prolapse. A bacterial pathogen was isolated from the diseased bullfrog intestines. The bacterium was identified as Vibrio cholerae using morphological, biochemical and 16S rRNA phylogenetic analysis. In this study, V. cholerae was isolated and identified from diseased bullfrogs for the first time, providing a basis for the diagnosis and control of the disease. At the same time, it was also found that V. cholerae may be transmitted to humans from bullfrogs through bullfrog food and aquaculture water, creating a serious threat for human health. Therefore, society should pay attention to the modes of transmission of Vibrio cholerae from bullfrog and formulate reasonable safety measures to avoid disasters.

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Smooth move: Behavioural changes in captive western lowland gorillas (Gorilla gorilla gorilla) after group split and relocation

Konecna, M.; Marie, Z.; Cizkova, M.; Vodicka, R.; Slipogor, V.

2026-06-30 animal behavior and cognition 10.64898/2026.06.25.734437 medRxiv
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Zoological gardens strive to prioritize excellent animal care and adhere to the highest standards of animal welfare, ensuring that the conditions of animals` social and physical environment are as close as possible to those in the wild. This study investigates the effects of group split and relocation to the new enclosure on the behaviour of western lowland gorillas (Gorilla gorilla gorilla) (N = 7) living in Prague Zoo, Czech Republic. We conducted over 289 hours of behavioural observations focusing on daily activities, social interactions, and behaviours that serve as potential stress and welfare indicators. The group split led to establishment of two groups in two separate enclosures; the old enclosure consisted of a "bachelor group" (i.e. three males) and a new enclosure consisted of a mixed-sex group (i.e. three females and one juvenile male). The behavioural comparisons across different study periods were conducted using linear mixed models (LMMs). The changes led to an increase in time spent moving, feeding, being in social proximity, and higher rates of approaches among the gorillas, as well as to a decrease in rates of self-directed and "undesirable" behaviors. Our findings indicate that the gorillas effectively adapted to the changes, most likely by relying on social support, to navigate new conditions. This study contributes to our understanding of how socio-cognitively complex species cope with necessary alterations in captive animal care programs. Furthermore, these observations may inform strategies to enhance the welfare of zoo-housed animals and to improve their captive care.

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Trends and Gaps on Philippine Scombrid Research: A bibliometric analysis

Gadut, J.-A.; Deocaris, C. C.; Alinsug, M. V.

2021-07-01 bioinformatics 10.1101/2021.06.30.450467 medRxiv
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Philippine scombrids have been among the top priorities in fisheries research due primarily to their economic value worldwide. Assessment of the number of studies under general themes (diversity, ecology, taxonomy and systematics, diseases and parasites, and conservation) provides essential information to evaluate trends and gaps of research. This study presents a bibliometric evaluation of the temporal trends from 2000-2019 on Philippine scombrid research. Seven out of nine tuna species have SREA values equal to or higher than 1 with Thunnus albacares being the most researched tuna species. Twelve out of thirteen non-tuna species have SREA values less than one, thus indicating low research effort. It was apparent that there were significant differences in the number of studies in each thematic area except in chemical analysis and diseases and parasites between scombrid groups where there was low research effort observed. The research points at the uneven research distributions between scombrid groups in each thematic area. As locally published research are significantly behind foreign publications in terms of citation index, international collaborations by Filipino researchers have shown an increase research impact. Our study hopes to influence the local and international R&D agenda on Philippine scombrids and promote solidarity among nations towards its conservation and management.

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Statistical invisibility of working equids in Mexico: Dissecting the gap between global diagnostics and official data (1970-2022).

Garcia-Seco, E.; Diaz, M. A.; Tadich Gallo, T.; Toribio, R. E.; Galindo Maldonado, F.; Hernandez-Gil, M.

2026-04-17 scientific communication and education 10.64898/2026.04.15.718791 medRxiv
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BackgroundWorking equids are fundamental to the socioeconomic structure of Mexicos small-scale agricultural sector, which accounts for 71.2% of the countrys active Agricultural Production Units (APUs). Despite their critical role in human rural livelihoods, food security, and sustainable development, these animals face systemic "statistical invisibility" within national and international productive frameworks. This study evaluates the long-term population dynamics and geographical distribution of working equids to analyze their current status amidst agricultural modernization. MethodsA retrospective analysis was conducted using national census data from 1970 to 2022 provided by the National Institute of Statistics, Geography, and Informatics (INEGI). Population trends for horses, donkeys, and mules were calculated using the Average Annual Variation Rate (AAVR). The severity of population declines was classified according to an adaptation of the International Union for Conservation of Nature (IUCN) criteria. Finally, national census records from INEGI, Agri-food and Fisheries Information Service (SIAP) and The Ministry of Agriculture and Rural Development (SADER) were contrasted with FAOSTAT database estimates to identify reporting discrepancies. ResultsBetween 1970 and 2022, the total equine population in Mexico decreased by 76.5%, falling from 6.8 to 1.6 million. However, a "paradox of modernization" was identified: while total numbers plummeted, the proportion of equids used specifically for work reached a historical peak of 81% in 2022, effectively having doubled from the 44% recorded in 2007. While donkeys and mules have suffered drastic total reductions (87% and 88%, respectively), working horses experienced a resilient 37% recovery between 2007 and 2022 (+3.71% AAVR). Furthermore, a staggering 710.8% discrepancy was found between national census data and FAOSTAT estimates, representing an overestimation of 11.3 million animals in international records. ConclusionsThe persistence and recent recovery of working equids reflect a "resilience of necessity" for approximately 500,000 APUs that depend exclusively on animal traction and packing due to economic constraints and complex topography. These findings challenge the narrative of total agricultural mechanization and highlight an urgent need for evidence-based public policies that address the statistical invisibility of working equids as indispensable drivers of rural sustainability and food security.

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Combined use of Candida Utilis and Idesia polycarpa var. Vestita Fruit Improve the Production Performance of Laying Quail

Liu, X.; Li, N.; Shu, Y.; Sun, Y.; Li, Y.; Hua, W.; Ye, Y.; Chen, F.; Tang, L.

2021-09-30 microbiology 10.1101/471201 medRxiv
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Although Idesia polycarpa oil has been wildly explored as a raw material for biodiesel, the reports studying the by-product Idesia polycarpa fruit residues (IPR) are few. This study aimed to evaluate the effect of the Idesia polycarpa fruit residues fermented feed additive (IPFF) on the egg production of laying quails. The egg production and related performances include egg quality, yolk cholesterol, yolk fatty acid, quails jejunum morphology, and relative gene expression were determined in this study. Compared to the standard diet group, birds fed the 1% IPFF showed a higher egg production (87.7% on average, 11.5% above the control; P<0.01). The yolk fatty acid composition and n6/n3 ratio were affected by IPFF or IPR. Compared to the standard diet group, the egg cholesterol content was lower in both IPFF and IPR groups, and the yolk n6/n3 ratio in the 5% IPFF group (10.3; P<0.01) was more reasonable. Meanwhile, birds under IPFF dietary supplement showed a thicker jejunum wall, higher villus, and deeper crypt than the standard diet group. In addition, the altered mRNA expression of four genes involved in cholesterol and fatty acids metabolism (SREBP-1, SREBP-2, ADGL, APOVLDL-II) in the 1% IPFF group and 5% IPR group indicated that the lipids metabolism and transportation were enhanced in the interclavicular fat pad and liver, relative to the standard diet group. HIGHLIGHTSEgg production was higher in IPFF groups Egg cholesterol was lower in IPFF groups Lipid metabolism and transportation was enhanced in IPFF groups Intestine wall was thicker in IPFF groups

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Prevalence and associated risk factors assessment of bovine fasciolosis in and around Bahir Dar, Ethiopia

Temesgen, A. B.; Mesfin, T.

2024-08-09 pathology 10.1101/2024.08.07.606800 medRxiv
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Cattle are among the most important livestock resources in Ethiopia, contributing significantly to the agricultural economy and rural livelihoods. They provide meat, milk, hides, draft power for crop production, and serve as a major source of income for farmers. Despite their vital role, cattle productivity is often constrained by various diseases, particularly parasitic diseases. One of the most significant of these is bovine fasciolosis, a condition caused by ingestion of metacercariae of liver flukes belonging to the genus Fasciola. This study aimed to assess the prevalence and associated risk factors of bovine fasciolosis in bahir dar, Ethiopia. A cross-sectional study was conducted from November 2021 to April 2022. A total of 384 cattle were randomly selected from different locations within the study area. Animals of all age groups and both sexes were included. Fecal samples were collected directly from the rectum of each animal using clean, labeled containers. The samples were examined using standard coprological techniques, specifically the sedimentation method, to detect liver fluke eggs. All findings were recorded, and the data were analyzed using descriptive statistical methods. The overall prevalence of fasciolosis was 49.21% (n=189). Based on origin, Sebatamit had the most incidence at 61.84% (n=47), followed by Kebele 11 at 59.37% (n=57), Tikurit at 50% (n=59), and Latammba at 27.65% (n=26). Statistical analysis revealed significant disparities in occurrence among areas. Cattle in poor condition had the largest prevalence (n=80, 64%), followed by medium condition (n=85, 50%) and fat cattle (n=24, 26.96%). This variation was statistically significant. Age-group analysis revealed comparable prevalence rates, with young cattle at 50.38% (n=65), adults at 47.33% (n=71), and elderly cattle at 50.47% (n=53), with no significant differences found. There were no significant sex-related variations in prevalence, with males exhibiting a prevalence of 49.73% (n=93) and females 48.73% (n=96). Local cattle had a slightly higher prevalence (n=111, 51.62%) than crossbreds (n=78, 46.15%), although the difference was not statistically significant (P=.29).These findings underscore the need for targeted, location-specific control strategies and highlight the importance of improved nutritional and health management practices to reduce the burden of fasciolosis in cattle populations.

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Seminal extracellular vesicles from boar AI doses contain fertility-predictive protein and miRNA cargo and improve sperm physiology

Martin-San Juan, A.; Cerrato Martin-Hinojal, C.; Nieto-Cristobal, H.; Martinez-Alborcia, M. J.; de Mercado, E.; Alvarez-Rodriguez, M.

2026-03-18 molecular biology 10.64898/2026.03.16.712050 medRxiv
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15.7%
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Boar semen contains spermatozoa and seminal plasma (SP) that carries extracellular vesicles (EVs) among other components. However, artificial insemination (AI) doses produced by AI companies are highly diluted based solely on sperm concentration. The aim of this study was to evaluate the integrity of EVs isolated from AI doses, characterize the protein and miRNA content from high-fertility (HF) and reduced-fertility (RF) boars, and evaluate their functional impact on spermatozoa after dilution by a coincubation up to 24 hours at 38 {degrees}C. Proteomics identified 108 differentially expressed proteins between HF and RF EVs (97 upregulated in HF, 11 in RF), and transcriptomics revealed 80 differentially expressed miRNAs (DEMs) in EVs, 52 in SP, and 3 in spermatozoa, showing inverse expression in various shared DEMs between fertility rates, suggesting compartment-specific regulation. Functional coincubation demonstrated that EVs remain biologically active after dilution. HF EVs improved sperm quality parameters and reduced oxidative stress, while RF EVs increased total and progressive motility. Overall, our findings show that EVs from AI doses retain structural integrity, carry fertility-associated protein and miRNA signatures, and functionally modulate sperm quality in vitro. These features highlight porcine EVs as promising biomarkers and potential tools to optimize reproductive performance in swine production.

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Internal and external features of wild birds after collisions without apparent trauma in Japan

Ushine, N.; Tanaka, A.; Sato, T.; Nonagase, M.; Hayama, S.-i.

2021-05-25 zoology 10.1101/2021.05.25.445584 medRxiv
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15.6%
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Wild birds often require rehabilitation after collisions, even with no apparent injury. Information about aftermath of collisions is still scares. Here, we investigated external characteristics and clinical features of the internal organs of wild birds that experienced collision and compared them with birds admitted to rehabilitation center for other reasons. Necropsy was performed on 55 bird carcasses from Passeriformes and Coraciiform. Five external characteristics were recorded before necropsy including; cause of admission, keel score, life stage, fat score, and number of days before death. The median survival time was calculated by Kaplan-Meier estimates. Data on external and internal features were compared using univariate and multivariate multinomial regressions. There was no significant difference in the median survival time among the causes of admission: 1 day for collision, 2 days for trauma, and 2 days for malnutrition. Kidney discoloration was more significantly associated with collision than with other trauma (p = 0.01). Although no apparent kidney abnormality (including enlargement) were observed, anterior lobe was significantly larger than posterior lobe with collision compared with malnutrition (p = 0.045). Birds that experienced collision exhibited a higher fat score than malnourished birds (p = 0.03). Our results suggested that wild birds with abundant fat were more likely to be admitted due to collision. The gross characteristics of collision included kidney discoloration and anterior lobe extension, which was possibly due to rupture of renal blood vessels by blunt external force. From these findings, it was considered that collision caused major axis of anterior lobe significantly larger than posterior lobe, even though no abnormal finding in renal size such as hypertrophy was recognized. This was the first study to evaluate the cause of admission, necropsy results, and external characteristics in wild birds admitted to rehabilitation centers. Absolute cage rest should be adhered to restore renal function for those birds admitted due to collision, and handling and treatment should be minimized to avoid excess movement of the birds.

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Facial expressions of emotional stress in horses

Lundblad, J.; Rashid, M.; Rhodin, M.; Haubro Andersen, P.

2020-10-19 animal behavior and cognition 10.1101/2020.10.19.345231 medRxiv
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15.5%
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Horses have the ability to generate a remarkable repertoire of facial expressions, some which have been linked to certain emotional states, for example pain. Studies suggest that facial expressions may be a more honest expression of emotional state in horses than behavioral or physiological parameters. This study sought to describe the facial expressions during stress of healthy horses free of pain, using a standardized method of recording facial expressions in video. Stress was induced in 28 horses by subjecting them to road transport and 10 of these horses were also subjected to social isolation. The horses served as their own control. A body-mounted, remote controlled heart rate monitor provided continuous heart rate measurements during the interventions. The horses facial expressions were video-recorded during the interventions. Frequency and duration of each facial expression were then determined, according to the Equine Facial Action Coding System. Heart rate increased during the stressful interventions (p=0.01), confirming that the interventions were stressful. Using both the human investigation- and the co-occurrence methods, the following facial traits could be observed during stress: eye white increase (p<0.001), nostril dilator (p<0.001), upper eyelid raiser (p<0.001), inner brow raiser (p=0.042), tongue show (p<0.001) along with an increase in ear flicker (p<0.001) and blink frequency (p<0.001). The facial actions were successfully used to train a machine-learning classifier to discriminate between stressed and calm horses, with an accuracy of 74.2 %. Most of the facial features identified correspond well with previous research on the subject, for example flared nostrils, repetitive mouth behaviors, increased eye white, tongue show and ear movements. Some features selected as indicative of emotional pain-free stress are used in face-based pain assessment tools, such as dilated nostrils, eye white increase or inner brow raiser. The relation between facial expressions of stress and pain should therefore further be studied.

20
Hybrid yak-cattle in situ conservation via interspecies somatic cell nuclear transfer at ultra-high-altitude region

Yu, D.; Nima, Q.; Cao, L.; Gu, S.; Yin, K.; Zhang, Y.; Wang, J.; Basang, W.; Huang, Y.

2025-07-16 zoology 10.1101/2025.07.10.664179 medRxiv
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Pien-niu, the hybrid offspring of yak (Bos grunniens) and domestic cattle (Bos taurus), possess exceptional adaptability to ultra-high-altitude environments of the Qinghai-Xizang Plateau. In situ conservation is of special significance for ultra-high-altitude region because the offsprings need to possess the ability to adapt to the unique environment. The present study was to clone Pien-niu in Xizang via interspecies somatic cell nuclear transfer (iSCNT) to practice in situ conservation of large animal at ultra-high-altitude region. Ear fibroblast cells were isolated from Pien-niu at Qushui Experimental Station (3650m above sea level), an animal live conservation farm at Xizang, and used as iSCNT donor cells. The iSCNT blastocysts were transferred into the oviduct of surrogate Pien-niu which are maintained at Qushui Experimental Station. A live cloned Pien-niu calf was born on May 12, 2025 and keeps healthy till now. Results of short tandem repeat analysis revealed that the microsatellite loci of cloned calf were completely matched with that of donor fibroblast cells. The successful cloning of Pien-niu can provide a paradigm for in situ germplasm conservation via iSCNT at ultra-high-altitude region.