Analysis of current methods and Welfare concerns in the transport of 118 horses by commercial air cargo companies.

BACKGROUND: Studies on equine air transport practices and consequences are scarce. This prospective study aimed to describe horse and air journey details and practices, document how horse behavior and health changed during the air transport phases, quantify the occurrence of welfare issues, and identify possible associations between horse and journey details, air transport practices, and welfare issues. RESULTS: Data were collected from before departure to five days after arrival on 118/597 horses traveling on 32 commercial air journeys on different routes, varying in duration and conditions. Most horses were middle-aged warmblood females, 26% of which were pregnant, and being moved by air for sales. Before flying, most were quarantined (median: 18; IQR: 9-53 days), and their fitness for travel was certified by veterinarians. At the departure airports, external temperatures varied from - 6 °C to 33 °C, and horses were loaded by experienced flight grooms (median: 35; IQR: 15-40 years) into jet stalls (three-horse: 87%, two-horse: 13%). During the flights, horses were regularly watered (water intake median: 14 L) and fed ad libitum (feed consumption median: 8 kg). At the arrival airport, horses were unloaded from the jet stalls, and external temperatures ranged from - 5 °C to 32 °C. Then, all horses were transported to arrival quarantine by road. Air transport phases affected horses' health status and behavior; increased heart and respiratory rates and behaviors, such as pawing, head tossing, and vocalization, were mainly identified at departure and arrival. Horse interaction, nasal discharge, increased capillary refill time (CRT), and abnormal demeanor were observed more often one hour before landing while resting and normal capillary refill time were more often displayed five days after arrival (all P < 0.01). One hour before landing, horses with bad temperament and horses of unknown temperament were more likely to develop nasal discharge when transported in winter and autumn (P < 0.001). The likelihood of an increased CRT was associated with shorter flights in horses of unknown travel experience (P < 0.001). Ten horses were injured, and 11 developed pleuropneumonias (i.e., shipping fever). CONCLUSIONS: Air transport is a complex procedure with several different phases affecting horse health and behavior. Therefore, experienced staff should carefully manage each horse before, during, and after air journeys to minimize welfare hazards.

Dromedary camel's welfare: literature from 1980 to 2023 with a text mining and topic analysis approach.

Dromedary camels are the preferable livestock species in the arid and semi-arid regions of the world. Most of the world's camel populations are managed under a subsistence/extensive system maintained by migratory pastoralists but intensification is getting more frequent. Even though recently the welfare of camels has been receiving more attention, in many countries there are no regulations to protect their health and welfare. The objectives of this article were to explore the main research topics related to camel welfare, their distribution over time and to highlight research gaps. A literature search was performed to identify records published in English from January 1980 to March 2023 on Dromedary camel welfare via Scopus®, using "Camel welfare," "Camel behaviour," "She-camel" and "Camel management" as search words. A total of 234 records were retained for analysis after automatic and manual screening procedures. Descriptive statistics, text mining (TM) and topic analysis (TA) were performed. The result shows that even though there were fluctuations between years, records on camel welfare have increased exponentially over time. Asia was the region where most of the corresponding authors were located. The first five most frequent words were, "milk," "calv," "behaviour," "femal," and "breed," the least frequent word was "stabl." TA resulted in the five most relevant topics dealing with "Calf management and milk production," "Camel health and management system," "Female and male reproduction," "Camel behaviour and feeding," and "Camel welfare." The topics that contained the oldest records were "female and male reproduction" and "camel health and management system" (in 1980 and 1983, respectively), while the topic named "camel behaviour and feeding" had the first article published in 2000. Overall, even though topics related to camel behaviour and welfare are receiving more attention from academia, research is still needed to fully understand how to safeguard welfare in Dromedary camels.

Factors associated with house-soiling in Italian cats.

OBJECTIVES: The aim of the present study was to identify factors associated with house-soiling in Italian cats. METHODS: A cross-sectional online survey collected information on respondents' and cats' details and litter management, and whether the cat showed house-soiling. Univariable and multivariable regression models were performed using house-soiling (present/absent) and the type of house-soiling (ie, urinary, faecal, concurrent urinary and faecal) as dummy variables. RESULTS: Data from 3106 cats were obtained. The number of dogs and other cats in the household, the cat's age, the number, type and location of the litter boxes, the type of litter, and the frequency of litter scooping and full replacement were retained in the final multivariable regression model for house-soiling (model P <0.001, Akaike information criterion [AIC] = 2454.30). Urinary tract diseases, the type and number of litter boxes and the number of dogs in the household were associated with urinary house-soiling (model P <0.001, AIC = 534.08), and gastroenteric/musculoskeletal diseases, number of litter boxes and litter box location were associated with faecal house-soiling (model P <0.001, AIC = 448.52). Healthy cats, the number of dogs in the household, the type of litter and litter full replacement frequency were retained in the final multivariable regression model for the concurrent expression of urinary and faecal house-soiling (model P <0.001, AIC = 411.47). CONCLUSIONS AND RELEVANCE: Meeting cats' preferences for litter and litter box type, location, behavioural needs and strict litter hygienic conditions is recommended. Cat owners need to be educated to prevent and manage house-soiling in their cats.

Cats' and dogs' welfare: text mining and topics modeling analysis of the scientific literature.

Animal welfare is a field with increasing significance and has been raising huge concerns of the public and the political stage. Cats and dogs possess an important role in human life, but their welfare is not always secured from a legal aspect. This review aimed to describe the evolution and geographical distribution of "cats and dogs" and "puppies and kittens" welfare literature over the last 40 years, distinguish the main research topics studied and highlight gaps in knowledge. A search using Scopus® was performed with different search strings and predetermined filters as time range, language, and subject area. A total of 2,725 scientific literature records were retrieved but only the ones that referred to cats and dogs' welfare aspects were retained. The final 1,775 records were processed through descriptive statistics, and text mining and topic analysis procedures were performed on their titles and abstracts. The results showed that the number of studies has been increasing, especially in Europe and North America. "Shelter" was the most frequent word, followed by "behavior," "owner" and "adopt." The nine topics that emerged from the analysis were breeding, stress and housing conditions, welfare and pain assessment, public health, shelter management and euthanasia, behavioral problems, health issues and management, human-animal interaction, and owners' and veterinarians' perceptions. While stress and housing conditions, public health, and owners' and veterinarians' perceptions were the most studied topics, human-animal interaction was the least studied. This review confirmed the increasing research and interest in cats' and dogs' welfare and showed gaps in knowledge where further studies are needed.

Heart and brain: Change in cardiac entropy is related to lateralised visual inspection in horses.

Cerebral lateralisation is the tendency for an individual to preferentially use one side of their brain and is apparent in the biased use of paired sensory organs. Horses vary in eye use when viewing a novel stimulus which may be due to different physiological reactions. To understand the interplay between physiology and lateralisation, we presented a novel object (an inflated balloon) to 20 horses while electrocardiogram traces were collected. We measured the amount of time each horse looked at the balloon with each eye. We calculated 'sample entropy' as a measure of non-linear heart rate variability both prior to and during the stimulus presentation. A smaller drop in sample entropy values between the habituation phase and the sample presentation indicates the maintenance of a more complex signal associated with a relaxed physiological state. Horses that spent longer viewing the balloon with their left eye had a greater reduction in sample entropy, while time spend looking with the right eye was unrelated to the change in sample entropy. Therefore, the horses that exhibited a greater reduction in sample entropy tended to use their right hemisphere more, which may take precedence in emotional reactions. These results may help to explain the variation in lateralisation observed among horses.

Investigating the effects of essential oils and pure botanical compounds against Eimeria tenella in vitro.

Essential oils (EO) and natural bioactive compounds are well-known antibacterial and anti-inflammatory factors; however, little is known about their anticoccidial activity and mode of action. EO deriving from basil (BEO), garlic (GAR), oregano (OEO), thyme (TEO), and their main bioactive compounds were investigated for their anticoccidial proprieties and compared to salinomycin (SAL) and amprolium (AMP) in vitro. The invasion of Eimeria tenella sporozoites was studied on 2 cell models: Madin-Darby Bovine Kidney (MDBK) cells and primary chicken epithelial cells (cIEC). Invasion efficiency was evaluated at 2 and 24 h postinfection (hpi) with counts of extracellular sporozoites and by detection of intracellular E. tenella DNA by PCR. Results show that at both timepoints, the EO were most effective in preventing the invasion of E. tenella with an average reduction of invasion at 24 hpi by 36% in cIEC and 55% in MDBK. The study also examined cytokine gene expression in cIEC at 24 hpi and found that AMP, BEO, OEO, TEO, carvacrol (CAR), and thymol (THY) significantly reduced interleukin (IL)8 expression, with CAR also reducing expression of IL1ß and IL6 compared to the infected control. In addition, this work investigated the morphology of E. tenella sporozoites treated with anticoccidial drugs and EO using a scanning electron microscope. All the treatments induced morphological anomalies, characterized by a reduction of area, perimeter and length of sporozoites. SAL had a significant impact on altering sporozoite shape only at 24 h, whereas CAR and THY significantly compromised the morphology already at 2 hpi, compared to the untreated control. OEO and GAR showed the most significant alterations among all the treatments. The findings of this study highlight the potential of EO as an alternative to traditional anticoccidial drugs in controlling E. tenella invasion and in modulating primary immune response.

Listeria monocytogenes Strains Persisting in a Meat Processing Plant in Central Italy: Use of Whole Genome Sequencing and In Vitro Adhesion and Invasion Assays to Decipher Their Virulence Potential.

In this study, we used both a WGS and an in vitro approach to study the virulence potential of nine Listeria monocytogenes (Lm) strains belonging to genetic clusters persisting in a meat processing plant in Central Italy. The studied clusters belonged to CC1-ST1, CC9-ST9, and CC218-ST2801. All the CC1 and CC218 strains presented the same accessory virulence genes (LIPI-3, gltA, gltB, and aut_IVb). CC1 and CC9 strains presented a gene profile similarity of 22.6% as well as CC9 and CC218 isolates. CC1 and CC218 showed a similarity of 45.2% of the same virulence profile. The hypervirulent strains of lineage I (CC1 and CC218) presented a greater ability to adhere and invade Caco-2 cells than hypovirulent ones (CC9). CC1 strains were significantly more adhesive and invasive compared with CC9 and CC218 strains, although these last CCs presented the same accessory virulence genes. No statistically significant difference was found comparing CC218 with CC9 strains. This study provided for the first time data on the in vitro adhesiveness and invasiveness of CC218-ST2801 and added more data on the virulence characteristics of CC1 and CC9. What we observed confirmed that the ability of Lm to adhere to and invade human cells in vitro is not always decipherable from its virulence gene profile.

Litter Management Practices and House-Soiling in Italian Cats.

There are about 10.1 million domestic cats in Italy, but information on cats' litter management and house-soiling prevalence is scant. This study described cats' and cat owners' profiles, litter management practices, and whether cats show house-soiling, also comparing between professionals (i.e., breeders) and amateurs (i.e., pet owners). A cross-sectional online survey sought respondents' housing, family, and cat details, as well as other pet details, litter details, and whether the cats showed house-soiling. Data for a total of 3106 cats were obtained. Italian cats lived mainly in apartments, along with other cats or dogs. Italians owned mostly adult European breed cats, to whom they provided covered litter boxes filled with clumping substrates, scooped daily, and completely replaced weekly. Litter cleaning was more frequent when cats were owned for financial purposes (i.e., breeders) rather than for companionship, but more space was provided for pets than for breeding cats. The recalled prevalence of elimination problems (16.7%) was lower compared to other studies, with cats mainly eliminating urine (54.6%) on objects in squatting posture (35.2%). Overall, this research increased our understanding of cat litter management in Italy. These findings could fill a gap in the knowledge regarding litter management and house-soiling incidences in Italy. Further studies to investigate possible risk factors for house-soiling are needed.

Establishment and characterization of an SV40 immortalized chicken intestinal epithelial cell line.

Primary chicken intestinal epithelial cells or 3D enteroids are a powerful tool to study the different biological mechanisms that occur in the chicken intestine. Unfortunately, they are not ideal for large-scale screening or long-term studies due to their short lifespan. Moreover, they require expensive culture media, coatings, or the usage of live embryos for each isolation. The aim of this study was to establish and characterize an immortalized chicken intestinal epithelial cell line to help the study of host-pathogen interactions in poultry. This cell line was established by transducing into primary chicken enterocytes the SV40 large-T antigen through a lentiviral vector. The transduced cells grew without changes up to 40 passages maintaining, after a differentiation phase of 48 h with epidermal growth factor, the biological properties of mature enterocytes such as alkaline phosphatase activity and tight junction formation. Immortalized enterocytes were able to generate a cytokine response during an inflammatory challenge, and showed to be susceptible to Eimeria tenella sporozoites invasion and generate a proper immune response to parasitic and lipopolysaccharide (Escherichia coli) stimulation. This immortalized cell line could be a cost-effective and easy-to-maintain model for all the public health, food safety, or research and pharmaceutical laboratories that study host-pathogen interactions, foodborne pathogens, and food or feed science in vitro.

Detour test performance of cloned minipigs from three different clone populations.

Genetics, the uterine environment, maternal behavior, and rearing conditions can all influence animal behavioral phenotypes. Some studies on cloned pigs have found no differences between the behavioral patterns of cloned and non-cloned animals. Other studies conducted on dogs have reported similarities in the behavior of cloned subjects. This study evaluated the performance of 12 cloned minipigs from three different clone populations (A, B, C) in a detour test around symmetric and asymmetric barriers. We measured the detour time and patterns, in order to investigate the pigs' cognitive abilities.The detour time and the detour entry/exit pattern were recorded. All the animals tended to keep a fixed entry/exit pattern instead of modifying it to accommodate changes in the working set. Significant differences in detour time were found among the populations, with animals belonging to population B being faster than the others, and also within each population.Our study is one of the few to assess the cognitive abilities of cloned minipigs. The results indicate that even animals belonging to the same cloned population may develop different cognitive, hence behavioral characteristics. Whether cloning can be utilized to obtain similar behavioral phenotypes therefore remains a matter of debate.

GLAD Scale for Ranking Welfare of Horses on Arrival after Transport to Slaughterhouses.

To date, there is no official method for measuring horse welfare after transport. This study aimed to develop a scale to classify horses into four categories: good shape; light affected; affected; down (GLAD) based on their welfare impairment measured at unloading. To this end, 15 animal-based measures (ABMs), previously recorded from 1019 horses, were scored. Weight and severity scores provided by welfare experts, alongside the number of welfare principles highlighted by the ABM, were assigned to each ABM. The welfare impairment (S) of each horse was then calculated as the weighted sum of the severity scores of the 15 ABMs. Three thresholds were also set to define the four GLAD categories; the ABM "down" (i.e., horses unable to stand and walk on arrival, also considered by the law as the indicator of the worst welfare) was used as the higher threshold, Sdown, (category D); the intermediate threshold, S2, was defined by the ABM "injuries", assumed to represent highly impaired welfare (category A); the threshold, S1, was defined assuming that significant welfare impairment starts from 20% of S2 (L category). Horses with an S value below S1 were considered physically and mentally fit (G category). Out of 1019, 43% of horses fell into category G, 48% into L, 9% into A, and 0.3% into D. Our scale could be useful for veterinarians to decide whether a horse can be slaughtered immediately (G), needs rest (L), needs attention (A), or euthanasia (D), but further validation is needed.

Autonomic nervous system balance in parturient mares: Spontaneous vs induced delivery.

Delivery is not easily predictable in horses and the consequences of dystocia can be serious for both the mare and foal. An induction protocol with low doses of oxytocin has been reported as a safe procedure. This study investigates the effect of induced delivery on at-term mares' sympathetic-vagal balance. Fourteen mares were included and divided into two groups, one subjected to spontaneous delivery (SD), and one to induced delivery (ID). In both groups, an ECG was recorded using an elastic belt with integrated smart textile electrodes. The recording started before the delivery (Basal), continued close to delivery (Pre-delivery) and during delivery (Delivery), and ended after parturition (Placental expulsion). From the ECGs, Heart Rate Variability (HRV) parameters relating to time and frequency domains and non-linear analysis were extrapolated. The HRV analysis was performed both within the same group (IntraGA) and between the two groups (InterGA). In the present study, spontaneous and induced delivery did not appear to differ in autonomic nervous system functioning. In IntraGA analysis, both for SD and ID mares, delivery and placental expulsion periods were parasympathetic dominated since vagal-related HRV parameters increased. Moreover, no differences were found in InterGA comparison between SD and ID mares, except for the pre-delivery period of ID mares, during which both branches of the autonomic nervous system were activated. These results are in line with the literature on parasympathetic dominance during parturition and no change in Heart Rate Variability following exogenous oxytocin administration in parturient mares.

Pilot Study on Annual Horse Movements by Air and the Possible Effect of the Covid-19 Pandemic.

There is a lack of information on the number of horses shipped globally by air annually, the purpose of air travel and the routes of their journeys. This pilot study aimed to collect retrospective data on the international movements of horses by air from 2018 to 2021, describe their routes, and identify the possible effects of the coronavirus SARS-CoV-2 (COVID-19) pandemic. Equine transport data was gathered from 7 of 15 international shipping companies (ISCs) and 5 of 8 airlines contacted by email. The seven ISCs performed a median of 10,401 horse movements annually, ranging from a few hundred to several thousand movements per company, most frequently in Europe (Western and Northern Europe), Middle East/Africa (Middle East, Southern Africa), Asia Pacific (Australia), and the Americas (North and South America). The five airlines performed a median of 10,656 horse movements annually, importing and exporting horses to and from Europe, North America, Australasia, and the Middle East. For all but one airline, the number of horse movements decreased in 2020. The number and journey characteristics of horses transported by air require further scientific studies focused on the epidemiological and welfare risks unique to this type of transport to enable the development and implementation of best practices and regulations based on objective evidence.

Thyme, Oregano, and Garlic Essential Oils and Their Main Active Compounds Influence Eimeria tenella Intracellular Development.

Coccidiosis poses a significant challenge in poultry production and is typically managed with ionophores and chemical anticoccidials. However, the emergence of drug resistance and limitations on their use have encouraged the exploration of alternative solutions, including botanical compounds and improvements in in vitro screening methods. Prior research focused only on the impact of these alternatives on Eimeria invasion, with intracellular development in cell cultures receiving limited attention. This study assessed the impact of thyme (Thymus vulgaris), oregano (Origanum vulgare), and garlic (Allium sativum) essential oils, as well as their bioactive compounds, on the initial phase of schizogony in Madin-Darby bovine kidney cells, comparing their effectiveness to two commercially used anticoccidial drugs. Using image analysis and quantitative PCR, the study confirmed the efficacy of commercial anticoccidials in reducing invasion and schizont formation, and it found that essential oils were equally effective. Notably, thymol and carvacrol exhibited mild inhibition of intracellular replication of the parasite but significantly reduced schizont numbers, implying a potential reduction in pathogenicity. In conclusion, this research highlights the promise of essential oils and their bioactive components as viable alternatives to traditional anticoccidial drugs for mitigating coccidiosis in poultry, particularly by disrupting the intracellular development of the parasites.

Rider Variables Affecting the Stirrup Directional Force Asymmetry during Simulated Riding Trot.

Riders' asymmetry may cause back pain in both human and equine athletes. This pilot study aimed at documenting in a simple and quick way asymmetry in riders during a simulation of three different riding positions on wooden horseback using load cells applied on the stirrup leathers and identifying possible associations between riders' asymmetry and their gender, age, level of riding ability, years of riding experience, riding style, motivation of riding, primary discipline and handedness. After completing an interview to obtain the previously mentioned information, 147 riders performed a standardized test on a saddle fixed on a wooden horseback-shaped model. The riding simulation was split into three phases of 1 min each: (1) sit in the saddle, (2) standing in the stirrups and (3) rising trot. The directional force on the left and the right stirrup leathers was recorded every 0.2 s. A paired t-test was performed on the recorded data to test the difference (i.e., asymmetry) in each phase. In phases 1, 2 and 3, 99.3% (53.4% heavier on the right (R)), 98% (52.8% heavier on the left (L)) and 46.3% (51.5% heavier on the left (L)) of the riders were asymmetrical, respectively. Chi-square tests showed a significant association between riding ability and riding experience, but no significant association between reported handedness and calculated leg-sidedness (p > 0.05). Univariate logistic (1: asymmetry, 0: symmetry) regression analysis was performed only on the phase 3 data. One-hand riders were found twice more likely to be asymmetrical than two-hand riders (Odds Ratio (OR): 2.18, Confidence Interval (CI): 1.1−4.29; p = 0.024). This preliminary study confirmed that the majority of the riders are asymmetrical in load distribution on stirrups and suggested the riding style as a possible risk factor for asymmetry.

Journeys, Journey Conditions, and Welfare Assessment of Broken (Handled) Horses on Arrival at Italian Slaughterhouses.

During horse transportation, the journey conditions are considered a welfare risk. This study aimed to document journeys, journey conditions, and welfare status of handled horses on arrival at two different slaughterhouses in Northern and Southern Italy, to find possible associations between journey conditions and welfare problems. The welfare status of 613 draft-breed and light-breed horses from 32 different journeys was evaluated on arrival at the slaughterhouses with a standardized protocol, using animal-based (ABMs) and environmental-based (EBMs) measures. The drivers' skills and vehicle characteristics were found to be mostly compliant with EC 1/2005. The horses traveled in single bays, 90° to the direction of travel for an average journey duration of 26.5 ± 14 h. On arrival at the slaughterhouses, the horses were unloaded by handlers, via halter and rope. The prevalence of reluctance to unload, injuries, nasal, and lacrimal discharge was 22.2%, 24.6%, 11.6%, and 10%, respectively. Journey duration, unloading duration, vehicle changes, long stops, handlers/drivers' skills, temperature, season, and horse individual characteristics were associated with horses' welfare and health status (all p < 0.05). Our study confirms the hypothesis that appropriate journey conditions are of crucial importance to safeguard the welfare of broken/handled horses transported over long distances for slaughter.

A mixture of organic acids and thymol protects primary chicken intestinal epithelial cells from Clostridium perfringens infection in vitro.

Necrotic enteritis causes economic losses estimated to be up to 6 billion US dollars per year. Clinical and subclinical infections in poultry are also both correlated with decreased growth and feed efficiency. Moreover, in a context of increased antibiotic resistance, feed additives with enhanced antimicrobial properties are a useful and increasingly needed strategy. In this study, the protective effects of a blend of thymol and organic acids against the effects of Clostridium perfringens type A (CP) on chicken intestinal epithelial cells were investigated and compared to bacitracin, a widely used antibiotic in poultry production. Primary chicken intestinal epithelial cells were challenged with CP for a total time of 3 h to assess the beneficial effect of 2 doses of citric acid, dodecanoic acid, and thymol-containing blend, and compare them with bacitracin. During the challenge, different parameters were recorded, such as transepithelial electrical resistance, cell viability, mRNA expression, and reactive oxygen species production. CP induced inflammation with cytokine production and loss of epithelial barrier integrity. It was also able to induce reactive oxygen species production and increase the caspase expression leading to cellular death. The high dose of the blend acted similarly to bacitracin, preventing the disruptive effects of CP and inducing also an increase in zonula occludens-1 mRNA expression. The low dose only partially prevented the disruptive effects of CP but successfully reduced the associated inflammation. This study shows that the usage of thymol combined with 2 organic acids can protect primary chicken intestinal epithelial cells from CP-induced damages creating a valid candidate to substitute or adjuvate the antibiotic treatment against necrotic enteritis.

Journeys, Journey Conditions, and Welfare Assessment of Unbroken (Unhandled) Horses on Arrival at a Slaughterhouse in Italy.

Transportation of horses to slaughterhouses can pose a welfare concern, in particular when horses are unbroken/unhandled. This study aimed to describe their journeys and journey conditions, document their welfare status on arrival in a slaughterhouse in Italy, and investigate possible associations between journey conditions and welfare issues. A total of 395 unbroken draft horses in 20 different consignments were assessed with a standardized protocol. The most common departure point (16/20 consignments, 80%) was a French assembly center, but many of these horses had Spanish passports, suggesting they had previously been transported from Spain to France. The average journey duration was 34 ± 14 h, including journey breaks (i.e., short stops inside the vehicle and long resting stops at control posts), while transit time was 24 ± 4 h. The drivers were well experienced, the trucks were well equipped (i.e., forced ventilation, drinkers), and the horses traveled loose in small groups (n ≤ 4 horses) within pens inside the vehicle. On arrival at the slaughterhouse, the horses self-unloaded and showed minimal behavioral and health problems. The prevalence of injuries, diarrhea/abnormal feces, and nasal discharge were 1.52%, 6.58%, and 4.30%, respectively. Cold temperatures, space allowance, and lack of feeding during transport were found to be the main hazards of those problems (all p < 0.05). Our findings confirm that the correct assessment of the fitness for transport, adequate journey conditions, and experienced staff are crucial factors to safeguard the welfare of unbroken horses during long journeys.

Horses show individual level lateralisation when inspecting an unfamiliar and unexpected stimulus.

Animals must attend to a diverse array of stimuli in their environments. The emotional valence and salience of a stimulus can affect how this information is processed in the brain. Many species preferentially attend to negatively valent stimuli using the sensory organs on the left side of their body and hence the right hemisphere of their brain. Here, we investigated the lateralisation of visual attention to the rapid appearance of a stimulus (an inflated balloon) designed to induce an avoidance reaction and a negatively valent emotional state in 77 Italian saddle horses. Horses' eyes are laterally positioned on the head, and each eye projects primarily to the contralateral hemisphere, allowing eye use to be a proxy for preferential processing in one hemisphere of the brain. We predicted that horses would inspect the novel and unexpected stimulus with their left eye and hence right hemisphere. We found that horses primarily inspected the balloon with one eye, and most horses had a preferred eye to do so, however, we did not find a population level tendency for this to be the left or the right eye. The strength of this preference tended to decrease over time, with the horses using their non-preferred eye to inspect the balloon increasingly as the trial progressed. Our results confirm a lateralised eye use tendency when viewing negatively emotionally valent stimuli in horses, in agreement with previous findings. However, there was not any alignment of lateralisation at the group level in our sample, suggesting that the expression of lateralisation in horses depends on the sample population and testing context.

In Vitro Assessment of Anticoccidials: Methods and Molecules.

Avian coccidiosis is a disease causing considerable economic losses in the poultry industry. It is caused by Eimeria spp., protozoan parasites characterized by an exogenous-endogenous lifecycle. In vitro research on these pathogens is very complicated and lacks standardization. This review provides a description of the main in vitro protocols so far assessed focusing on the exogenous phase, with oocyst viability and sporulation assays, and on the endogenous phase, with invasion and developmental assays in cell cultures and in ovo. An overview of these in vitro applications to screen both old and new remedies and to understand the relative mode of action is also discussed.