Tympanic ossicles and pharyngeal arches

We have performed a study on 11 human embryos regarding the development of the tympanic ossicles and their relationship with the first pharyngeal arch. After performing measurements to date the embryos and foetuses chronologically, we performed a meticulous dissection of the temporal bones. Subsequently, they were fixed in 10% formol, decalcified with 2% nitric acid, embedded in Paraplast, sectioned in 7-mm sequences and stained with Martin's trichrome technique. In the 21- and 24-mm cranium-raquis (CR) length human embryos, we have observed the head of the malleus and the body of the incus close to Meckel's cartilage, in addition to the handle of the malleus, the long limb of the incus and the stapes. Between them there was a mesenchymal band inside the primordium of the tympanic cavity. In the 27-mm CR embryo, the various components of the malleus and incus were fusing, and in the 30-mm CR embryo the union was complete. From our observations, we can conclude that the malleus and the incus are derived from the first and second pharyngeal arches.     

A detailed subgross morphometric study on the auditory ossicles of the New Zealand rabbit

This study was designed to reveal, in detail, the features of the auditory ossicles of the New Zealand rabbit, one of the most frequently used animals in scientific researches. Heads of 10 dead adult New Zealand rabbits of both sexes weighing 2.5 +/- 0.2 kg were used in the study. The auditory ossicles were gently removed from their situs and observed. The auditory ossicles were three small bones: the malleus, the incus and the stapes. The lenticular bone seemed to be the lenticular process, as is in human beings. In general, macroanatomic features of the bones were similar to those reported in the literature. Findings in this study, however, differed greatly, especially in detail. There were statistically significant differences in the morphometric data of the features of the auditory ossicles between the right- and left-hand sides. Data from the right-hand side were significantly higher.     

VIRTUAL CT OTOSCOPY OF THE MIDDLE EAR AND OSSICLES IN DOGS

Virtual otoscopy enables noninvasive 3D endoluminal imaging of the middle ear through postprocessing of computed tompgraphy (CT) data. A standardized imaging approach for the middle ear was established in six normal dogs in an attempt to optimize the clinical application and student education. High‐resolution CT data were obtained. Virtual otoscopic images of the middle ear cavity and ossicles were generated using commercially available software. The views of the four different directions (the ear canal, tympanic bulla, eustachian tube, and ossicular chain) were made for virtual otoscopy. The promontory, cochlea window, tympanic bulla, septum bulla, and auditory tube were distinguished easily and clearly. One of the ossicles, the malleus, was visualized accurately. However, small structures such as the incus and stapes always could not be seen. The main advantage of virtual otoscopy is not only to provide diagnostic information but also to enhance the quality and efficiency of student education, because it contributes to an understanding of the anatomy of the middle ear. We describe the normal topographical 3Dimages of the middle ear of the dog using virtual otoscopy.     

Three-Dimensional Anatomy of the Temporal Bone in Normal Mice

This study was performed to determine the three-dimensional (3D) structure of the murine temporal bone and to provide a survey atlas of the temporal bone structures in mice. The temporal bones of adult BALB/c mice were examined and 3D high-resolution reconstructions of the temporal bone were obtained using a micro-CT system. Using the system described here, the bony labyrinth and membranous labyrinth could be investigated in a non-destructive manner. The turning rate of the cochlea was two (human rate: two and a half). The shapes of the superior and posterior semicircular canals were more flexed than those in humans. The malleus manubrium was directed anteriorly and was thin and fan-shaped like a Persian sword. The size of the incus relative to the malleus was smaller than that in the human ossicles. The 3D reconstruction of murine temporal bone described in this study provides anatomical information that will be useful in future studies using mouse model.     

Prenatal development in greater mouse‐eared bat,Myotis myotis (Borkhausen, 1797) (Chiroptera, Vespertilionidae)

Order Chiroptera is the second largest mammal group after rodents. An understanding of the development of the bats, which is a very special mammal group in terms of their lifestyles, morphology and their ability to fly, is very important because most of the adult anatomical differences characterizing species occur during organogenesis. In this study, developmental stages were determined for Myotis myotis species based on external morphological characteristics from embryos obtained from wild‐caught pregnant females. The developmental stages of M. myotis were comparable with those of other bat species.     

Uterus duplex bicollis,Vagina simplex in laboratory Guinea pigs (Cavia porcellus), rats (Rattus norvegicus forma domestica) and mice (Mus musculus forma domestica)

The female genital tract originates from the Müllerian ducts during embryological development. Fusion of the ducts occurs in different segments depending on the animal species, resulting in a variational number of the respective organ. Current literature on genital tract morphology of laboratory rodents is controversial. Therefore, the present study aimed at determining an anatomically correct definition of the uterus in laboratory guinea pigs, mice and rats. In all three rodent species, we found two separate cervical canals that communicate with an individual uterus via discrete ostia uteri interna. The correct anatomical definition should therefore be uterus duplex bicollis, vagina simplex.     

Virtual otoscopy for evaluating the inner ear with a fluid-filled tympanic cavity in dogs

The feasibility of virtual otoscopy (VO) imaging was evaluated in five dogs with experimentally induced otitis media, two control dogs, and two canine patients with otitis media. VO images of the tympanic cavity and ossicles were generated with commercially available software using raw computed tomography (CT) data. Eight out of 10 ears inoculated with pathogen exhibited obvious clinical signs associated with otitis externa. CT images revealed soft tissue density material occupying the tympanic bulla compatible with otitis media in three dogs with experimentally induced otitis media and two patients. No remarkable features were observed on the radiographs. Four different VO views (ear canal, tympanic bulla, eustachian tube, and ossicular chain) were created. Promontory, cochlea window, tympanic, and septum bulla as well as ossicles were easily and clearly distinguished except for the incus and stapes of the clinical patients. VO images were not more suitable than images created with conventional CT for accurately diagnosing otitis media in this study. However, it appears that VO could be more feasible for assessing the complex structure of the inner ear in dogs with fluid-filled tympanic cavities since fluid accumulation within the tympanic bulla did not affect the evaluation of bony tissue in the middle ear on VO images.     

Bovine leptospirosis in abattoirs in Uganda: Molecular detection and risk of exposure among workers

Leptospirosis is a zoonotic bacterial disease reported worldwide. In Uganda, seropositivity has been reported in both humans and domesticated animals, including cattle. However, it remains unknown whether cattle are shedding leptospires and thus acting as potential source for human leptospirosis. We conducted this cross‐sectional study in two cattle abattoirs in Kampala, Uganda between June and July 2017. Kidney and urine samples from 500 cattle sourced from across the country were analysed by real‐time PCR to establish the prevalence of Leptospira‐positive cattle and risk of exposure to abattoir workers. The species of infecting Leptospira was determined by amplification of secY gene and compared to reference sequences published in GenBank. Of 500 cattle tested, 36 (7.2%) had Leptospira DNA in their kidneys (carriers), 29 (5.8%) in their urine (shedders); with an overall prevalence (kidney and/or urine) of 8.8%. Leptospira borgpetersenii was confirmed as the infecting species in three cattle and Leptospira kirschneri in one animal. Male versus female cattle (OR = 3, p‐value 0.003), exotic versus local breeds (OR = 21.3, p‐value 0.002) or cattle from Western Uganda (OR = 4.4, p‐value 0.001) and from regions across the border (OR = 3.3, p‐value 0.032) versus from the central region were more likely to be Leptospira‐positive. The daily risk of exposure of abattoir workers to ≥1 (kidney and/or urine) positive carcass ranged from 27% (95% credibility interval 18.6–52.3) to 100% (95% CI 91.0–100.0), with halal butchers and pluck inspectors being at highest risk. In conclusion, cattle slaughtered at abattoirs in Uganda carry and shed pathogenic Leptospira species; and this may pose occupation‐related risk of exposure among workers in these abattoirs, with workers who handle larger numbers of animals being at higher risk.     

Functional Activity of Frozen Thawed Chinchilla lanigera Spermatozoa Cryopreserved with Glycerol or Ethylene Glycol

The cryopreservation of spermatozoa constitutes a valuable tool for the captive breeding management of valuable and/or threatened species. Chinchilla lanigera is a species almost extinct in the wild, and the domestic counterpart has one of the most valuable pelts in the world. The objectives of this study were to: (i) compare the functional activity of post‐thawed chinchilla spermatozoa cryopreserved at ?196°C either with glycerol (G) or ethylene glycol (EG) as cryoprotectants (1 m final concentration) and (ii) investigate the effects of incubating the gametes for 4 h in the presence or in the absence of the cryoprotectants; evaluations were performed taking into account motility, viability, response to hypo‐osmotic shock and acrosome integrity of the cells. Parameters reflecting post‐thaw (0 h) sperm functional activity were significantly lower than those of freshly ejaculated gametes. When comparing the cryoprotectant efficiency of G vs EG, neither cryoprotectant agent offered appreciable advantages. After 4 h of incubation, in the presence or absence of the cryoprotectant agent, a rapid and significant decrease was found in all functional parameters and remained at ～ 20–30% motile, viable and viable acrosome intact cells. Viability was significantly lower when the cryoprotectant was removed from the media (possibly due to the centrifugation process). With respect to the maintenance of sperm membrane integrity, only ～ 10% of cells showed membrane resistance to hypo‐osmotic conditions after the 4 h incubation period. These results constitute new insights for cryopreservation protocols and the development of assisted reproductive techniques in this species.     

Optimization of surfactin production from Bacillus subtilis in fermentation and its effects on Clostridium perfringens‐induced necrotic enteritis and growth performance in broilers

Bacillus species are commonly used as probiotics in the poultry feed industry for preventing infectious diseases and improving productivity by altering gastrointestinal microbiota. The growth parameters of Bacillus subtilis for surfactin production in fermentation and the benefits of surfactin on broiler chickens remain unclear. In this study, we examined the growth parameters of B. subtilis in fermentation and evaluated the effects of surfactin from B. subtilis‐fermented products on Clostridium perfringens‐induced necrotic enteritis and growth performance in broilers. Results showed that the highest viable biomass of B. subtilis was observed at 10% molasses and 2% yeast supplementation during fermentation. The 4‐ and 6‐day fermented B. subtilis products were heat‐, acid‐ and bile‐resistant. Furthermore, the 4‐day fermented B. subtilis products with the highest surfactin concentration showed the maximal antimicrobial activity against pathogens, including Escherichia coli, Staphylococcus aureus, Salmonella typhimurium and C. perfringens. Dietary B. subtilis‐fermented product supplementation in broilers significantly improved intestinal morphology and necrotic lesions under C. perfringens challenge. Bacillus subtilis treatments could enhance broiler productivity, as well as promote bone quality and intestinal morphology. These results together indicate that B. subtilis‐fermented products containing surfactin have potential for the development as feed additives and use as possible substitutes for antibiotics to treat C. perfringens in the poultry industry.     

Detection of hepatitis E virus RNA in rats caught in pig farms from Northern Italy

Hepatitis E virus (HEV) strains belonging to the Orthohepevirus genus are divided into four species (A–D). HEV strains included in the Orthohepevirus A species infect humans and several other mammals. Among them, the HEV‐3 and HEV‐4 genotypes are zoonotic and infect both humans and animals, of which, pigs and wild boar are the main reservoirs. Viruses belonging to the Orthohepevirus C species (HEV‐C) have been considered to infect rats of different species and carnivores. Recently, two studies reported the detection of HEV‐C1 (rat HEV) RNA in immunocompromised and immunocompetent patients, suggesting a possible transmission of rat HEV to humans. The role of rats and mice as reservoir of HEV and the potential zoonotic transmission is still poorly known and deserves further investigation. To this purpose, in this study, the presence of HEV RNA was investigated in the intestinal contents and liver samples from 47 Black rats (Rattus rattus) and 21 House mice (Mus musculus) captured in four pig farms in Northern Italy. The presence of both Orthohepevirus A and C was investigated by the real‐rime RT‐PCR specific for HEV‐1 to HEV‐4 genotypes of Orthohepevirus A species and by a broad spectrum hemi‐nested RT‐PCR capable of detecting different HEV species including rat HEV. The intestinal content from two Black rats resulted positive for HEV‐C1 RNA and for HEV‐3 RNA, respectively. None of the House mice was HEV RNA positive. Sequence analyses confirmed the detection of HEV‐C1, genotype G1 and HEV‐3 subtype e. The viral strain HEV‐3e detected in the rat was identical to swine HEV strains detected in the same farm. Liver samples were negative for the detection of either rat HEV or HEV‐3.     

Comparatively examining of the apelin‐13 levels in the Capoeta trutta (Heckel, 1843) and Cyprinus carpio (Linnaeus, 1758)

Apelin is a recently discovered peptide produced by several tissues in the various vertebrates and fish. Apelin has been suggested to have role in regulation of many diverse physiological functions including food intake, energy homoeostasis, immunity, osmoregulation and reproduction. In this study, apelin‐13 levels in the blood serum of Cyprinus carpio and Capoetta trutta were determined. Then the results were compared between two species and sexes of each species. Apelin‐13 level was analysed using the enzyme‐linked immunoassay (ELISA) kit (Rat apelin‐13 ELISA kit, catalog no: CSB‐E14367r). Apelin‐13 level in the blood serum of C. trutta was significantly higher than those of the C. carpio (p < 0.05). However, its levels were observed to be no significant difference (p > 0.05) that compared to between sexes of each species. There was a significant negative correlation (r = ?0.829, p = 0.0001) between the apelin‐13 level and body weight of C. carpio. However, no significant correlation (r = ?0.022, p = 0.924) between the apelin‐13 level and weight of C. trutta observed.     

Review of some pharmacokinetic and pharmacodynamic properties of anti‐infective medicines administered to the koala (Phascolarctos cinereus)

Although koalas are iconic Australian animals, no pharmacokinetic studies of any first‐line medicines used to treat diseased or injured koalas had been published prior to 2010. Traditionally, medicine dosages suggested for this species underwent linear extrapolation from those recommended for domesticated species. The koala, a specialist folivore whose natural diet consists of almost exclusively Eucalyptus spp. foliage has anatomical and physiological adaptations for detoxifying their diet which also affect medicine pharmacokinetic profiles. This review addresses aspects of medicine absorption, clearance, and other indices (such as medicine binding to plasma proteins) of enrofloxacin/marbofloxacin and chloramphenicol used for the systemic treatment of chlamydiosis, and fluconazole ± amphotericin, and posaconazole for the treatment of cryptococcosis. Based on observations from published studies, this review includes suggestions to improve therapeutic outcomes when administering medicines to diseased koalas.     

Seasonality and anatomical location of human tick bites in the United Kingdom

Tick bites on humans can occur in a variety of habitats and may result in the transmission of tick‐borne pathogens, such as the causative agent of Lyme borreliosis (LB), Borrelia burgdorferi sensu lato. As the risk of transmission of this pathogen to the host increases with the duration of tick feeding, the recognition and removal of ticks as soon as possible following attachment is important for reducing the risk of infection. Performing a thorough body examination for ticks following potential exposure is recommended by tick awareness campaigns. Knowledge of where on the body feeding ticks are frequently found, and at which times of year peak tick exposure occurs, provides important information for public health messaging and may aid those bitten by ticks to engage more effectively with tick‐checking behaviour. This paper summarizes human tick bites in the United Kingdom (UK) during 2013–2018 reported to Public Health England's passive Tick Surveillance Scheme and further examines the anatomical location and seasonality of bites from the most commonly encountered tick and LB vector Ixodes ricinus. A total of 1,328 tick records from humans were received of which 93% were I. ricinus. Humans were most commonly bitten by I. ricinus nymphs (70% bites). Tick bites were recorded on all parts of the body, but there were significant differences in their anatomical location on adults and children. Most tick bites on adults occurred on the legs (50%), whereas on children tick bites were mostly on the head and neck (43%). Bites from I. ricinus were recorded throughout the year but were most numerous during May to August. This study adds to the body of research on the seasonality and anatomical location of human tick bites in temperate Europe and highlights the importance of data collected through passive surveillance in addition to research and epidemiological studies.     

Lactobacillus ingluviei C37 from chicken inhibits inflammation in LPS‐stimulated mouse macrophages

Probiotics are growing alternatives to antibiotics, and can contribute to the prevention and treatment of diseases and enhance livestock production. Lactobacillus (L.) ingluviei is a novel probiotic species with growth‐enhancement effects; however, this species remains poorly understood, and there have been (to our knowledge) no studies focusing on its immunological effects. Here, we isolated L. ingluviei C37 (LIC37) from chicken and evaluated the bacterium's immunomodulatory properties to explore its probiotic potential. Real‐time quantitative PCR and ELISA showed that in vitro exposure of inflammation‐stimulated mouse peritoneal macrophages to heat‐killed LIC37 led to decreases in tumor necrosis factor‐α and interleukin (IL)‐6 levels and an increase in IL‐10. These findings suggested that LIC37 exerts anti‐inflammatory effects by modulating cytokine profiles. This species may be an attractive probiotic bacterial strain for use in animal production.     

Outbreak of swimmer's itch in Central Italy: Description,causative agent and preventive measures

Swimmer's itch is caused by the penetration of free‐swimming larvae of trematodes of the family Schistosomatidae in human skin. It is usually reported in people engaged in recreational water activities in freshwater bodies and in most of cases, it is provoked by bird schistosomes of the genus Trichobilharzia. In the summer 2017, many cases of dermatitis were recorded in people bathing in the waters of the Albano Lake (Rome, Italy) and a parasitological investigation was carried out in order to ascertain the causative agent of these cases. Snails of the family Lymnaeidae, natural intermediate hosts of bird schistosomes, were collected from lake shallow waters to detect the presence of trematodes of the genus Trichobilharzia. Pools of maximum 10 snails were placed in Petri dishes, and cercarial emergence was stimulated exposing snails to strong artificial light intensity at 25°C. Three hundred and thirty‐seven snails were collected and screened for the shedding of cercariae. Furcocercariae of the family Schistosomatidae, with a morphology overlapping that of the genus Trichobilharzia, were detected in seven Petri dishes. Assuming that in each positive Petri dish just one snail was shedding furcocercariae, the minimum infectious rate was 2.1%. Molecular analysis of furcocercariae allowed ascribing them to the species Trichobilharzia franki. Snails of the species Radix auricularia were identified as intermediate hosts of the parasite. This is the second record of T. franki causing cercarial dermatitis in Central Italy, the third in Italy. The 2017 was in Italy exceptionally warm and dry. Trematodes are sensitive to changes in temperature, being cercarial production and emission rates temperature dependent. Small increases in water temperature would speed up parasite development and transmission, leading to a manifold increase in cercarial emergence. Moreover, high temperatures raise chances to acquire the infection, due to increased time spent in water by people.     

Relation between cell‐bound exopolysaccharide production via plasmid‐encoded genes and rugose colony morphology in the probiotic Lactobacillus brevis KB290

The probiotic Lactobacillus brevis KB290 is a natural producer of cell‐bound exopolysaccharide (EPS), and the plasmid‐encoded glycosyltransferase genes are responsible for this EPS production. KB290 forms unique rugose colonies inside an agar medium; this characteristic is useful for detecting and enumerating KB290 in the gut or feces. However, the genetic elements associated with this morphology remain unclear. Here, we aimed to investigate the relation between the plasmid eps genes and rugose colony morphology in KB290. The plasmid‐cured mutants formed smooth colonies, and the rugose colony morphology was restored after complementation with the eps genes. The eps genes were successfully cloned and expressed in other L. brevis and L. plantarum strains. In these transformant strains, the presence of the EPS, consisting of glucose and N‐acetylglucosamine, correlated with rugose colonies, indicating that EPS is responsible for rugose colony formation. To the best of our knowledge, this is the first report identifying the genetic factors influencing rugose colonies in Lactobacillus strains. This rugose colony formation may serve as a useful selective marker for KB290 in routine laboratory and research settings and can be used to detect the spontaneous loss of plasmids in this strain.     

Hepatitis E virus in Common voles (Microtus arvalis) from an urban environment,Hungary: Discovery of a Cricetidae‐specific genotype of Orthohepevirus C

Hepatitis E virus is a major causative agent of acute hepatitis worldwide. Despite its zoonotic potential, there is limited information about the natural chain of hepevirus infection in wildlife, and the potential reservoir species. In this study, we performed a HEV survey by heminested RT‐PCR on rodent samples from an urban environment (in the city of Pécs, Hungary) and investigated the prevalence of the virus among these native rodent species (Apodemus agrarius, Apodemus flavicollis, Apodemus sylvaticus, Microtus arvalis and Myodes glareolus). HEV was detected exclusively in Common voles (M. arvalis), in 10.2% of screened voles, and 3.2% of all investigated samples from all species. Based on the phylogenetic analysis, our strain showed the closest homology with European Orthohepevirus C strains detected previously in faecal samples of birds of prey and Red fox, supporting the possibility of the dietary origin of these strains. In addition, our samples showed close phylogenetic relation with a South American strain detected in Necromys lasiurus (Cricetidae), but separated clearly from other Muridae‐associated strains, suggesting the presence of a Cricetidae‐specific genotype in Europe and South‐America. Based on these results, we hypothesize the reservoir role of M. arvalis rodents for the European Cricetidae‐specific Orthohepevirus C genotype.