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The use of non-penetrating pre-slaughter stunning (NPPSS) in ruminants remains a contentious issue in Indonesia since it is thought to interfere with the bleeding. This paper evaluated the bleeding time (BT), incidence of the false aneurysm (FA), and factors influencing these parameters in 460 NPPSS and 50 non-NPPSS slaughters in Indonesia. In this research, bodyweight, neck skin fold, sex, incisors condition, stunner strength, door closed-final stunning interval, number of shots, stunning success rate (SSR), stunning final-slaughter interval, broken skull, shooting placement, stunner-men and slaughtermen competencies, incision direction, incision location, BT, and FA were assessed for NPPSS slaughter. The same parameters were also assessed for non-NPPSS except the stunning parameter. The results showed that the BT of the non-NPPSS slaughters (187±47.37 seconds) was faster than that of the NPPSS slaughters, with the BT for the NPPSS dependent upon the level of unconsciousness. The longest BT was found in fully unconscious cattle shot once (304.32±69.76 seconds). The risk of FA incidence in non-NPPSS cattle was higher than in the NPPSS cattle. The FA incidence was 2.64 times higher in non-NPPSS cattle than in unconscious NPPSS cattle. The BT of non-NPPSS cattle was affected by FA, whereas the NPPSS cattle were affected by SSR, number of shots, broken skull, and FA incidence. The primary influence factor of FA in non-NPPSS cattle was incision location, whereas the NPPSS cattle were DCFS, incision location, and incision direction. Based on the results of this study, it can be concluded that the use of NPPSS prolongs BT and reduces the risk of FA in the slaughter of Australian Brahman cross cattle. [ABSTRACT FROM AUTHOR]

Skin injury at the ventral part of the body and joints of hind limbs are common in lesser mouse deer in captivity, especially when kept in unsuitable cages. These animals showed unstable behaviour due to stress that could lead to their inability to mount during mating. The recovery was long due to the constant in-contact of the lesions to the environment. However, recovery can be aided by antibiotic and supportive therapies, which include fluid and nutritional supplementations which can substitute severe fluid and protein losses. Broad spectrum of antibiotic should be used to prevent secondary bacterial infection. [ABSTRACT FROM AUTHOR]

Objective: We aimed to evaluate the potential of Ampiang-Dadih (AD), a combination of Indonesian traditional fermented buffalo milk ( Dadih ) and black glutinous rice flakes ( Ampiang ) as an anti-hypercholesterolemic agent and to prevent liver-cell degeneration using a rat model., Materials and Methods: A mixture of black glutinous rice powder (0.3 gm/gm feed) and fermented buffalo milk (3.74/200 gm BW) was prepared to obtain AD. Fifteen adult male Sprague Dawley rats were divided into three groups of five animals each: (A) negative control group (distilled water; 5 weeks), (B) hypercholesterolemia group (1% cholesterol per feed; 5 weeks), and (C) preventive AD group (1% cholesterol and AD; 5 weeks). The blood lipid profiles were measured at weeks 2, 4, and 5. The liver enzyme activity, cholesterol level, and histology were observed at the end of week 5., Results: AD administration simultaneously with cholesterol in Group C significantly prevented an increase in total plasma cholesterol and low-density lipoprotein levels compared to Group B. Alanine transaminase and aspartate transaminase were maintained at normal levels in Group C. Furthermore, the levels of liver cholesterol and liver cell degeneration in Group C were also maintained because of AD administration compared to that in Group B., Conclusion: This study demonstrated that AD has the potential to be developed as a functional food for hypercholesterolemia prevention., Competing Interests: The authors declare no conflict of interest., (© The authors.)

The fatty acid composition in the skin of Sunda porcupine (Hystrix javanica) is an interesting topic due to the special features of quills, especially in the dorsal region. Therefore, this study aims to analyze the composition of fatty acids in the dorsal region of Sunda porcupine skin. It was conducted using skin samples of the thoracodorsal and lumbosacral regions taken by biopsies and from frozen specimens. The skin lipid was extracted and then derivatized into fatty acid methyl ester before analyzing with gas chromatography mass spectrometry. The results showed that the skin is composed of up to 25 fatty acids ranging from C12 to C25 with various types but only 16 were found in both regions and sexes. Fatty acids with an antibacterial effect were found abundantly, such as oleic, palmitic, stearic, and linoleic acids. The total abundance in the thoracodorsal region was higher than lumbosacral, while the composition in male was higher than in female. Based on the results, the fatty acid composition in the dorsal skin region of Sunda porcupine consists of at least 16 types ranging from C12-C25. Additionally, the region and sex were observed to contribute significantly to the variation in skin fatty acid composition.

The complexity of the Sunda porcupine skin has become an important topic due to the unique characteristics of its quill follicles. The structure and chemical composition of the skin has affected many physiological and other conditions. Generally, quills are larger, stronger and stiffer than hair; therefore, the skin structure needs to adapt to support their physiology. The strength of the skin is determined by its collagen composition and arrangement; therefore, this study aims to analyse the composition and distribution of thick and thin fibres based on the specific characteristics of Sunda porcupine skin under polarized light using picrosirius red staining. The skin samples used were from the thoracodorsal and lumbosacral regions of eight Sunda porcupine adults. The histological staining was carried out using the picrosirius red method, while the samples were observed under a polarized light microscope and analysed with software. The results showed that the skin is composed of 36%-65% thick fibres, 20%-35% thin fibres and small amounts of other types with the lumbosacral region having higher compositions of thick and thin fibres than those in the thoracodorsal region. Furthermore, the thoracodorsal and lumbosacral regions have the highest composition of thick fibre in the deeper dermis and quill follicle, respectively. These demonstrated that the complexity of the skin structure of Sunda porcupine due to its quill derivates correlated with its collagen composition and distribution., (© 2022 Wiley-VCH GmbH.)

The Aceh cattle are local Indonesian beef cattle that are farmed in Aceh Province. This type of cattle is one of the sources of meat for the Aceh people. This study aims to analyze the quality of two primal cuts (longissimus lumborum and semitendinosus muscle) from Aceh cattle based on the muscle microstructure characteristics and MSTN gene expression. This study used a sample of longissimus lumborum and semitendinosus muscles from 18 adult male Aceh cattle with the age of 2-2.5 years and a BCS of 3.24. Muscle samples were obtained shortly after the cattle were slaughtered in slaughterhouses in Banda Aceh and Aceh Besar districts. Muscle microstructure analysis was performed using the HE, Masson's trichrome, and immunohistochemistry staining methods, while the MSTN gene expression analysis was performed using the qPCR method. The analysis of the physical quality of meat includes pH, meat color, fat color, cooking loss, water holding capacity, and WBSF value. The results showed that the area of LL muscle fibers was smaller than that of ST with relatively the same diameter. Both muscles were dominated by fast fibers with a percentage of 82.37% (LL muscle) and 91.80% (ST muscle). The area and composition of the type of muscle fibers are the main factors that influence the tenderness of Aceh beef. A higher distribution of collagen was found in ST muscles than in LL muscles. MSTN gene expression in both muscle types was relatively the same. Aceh cattle have large muscle fibers and are dominated by fast fibers with a high percentage, resulting in a low level of the tenderness of Aceh beef. However, the level of tenderness of Aceh beef is still in accordance with the cooking preparation of original and favorite cuisine of Aceh people., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2021 Hamny Sofyan et al.)

Here, we performed next-generation sequencing (NGS) on six large flying foxes (Pteropus vampyrus) collected in Indonesia. Seventy-five virus species in the liver tissue of each specimen were listed. Viral homologous sequences in the bat genome were identified from the listed viruses. This finding provides collateral evidence of viral endogenization into the host genome. We found that two of the six specimens bore partial sequences that were homologous to the plant pathogens Geminiviridae and Luteoviridae. These sequences were absent in the P. vampyrus chromosomal sequences. Hence, plant viral homologous sequences were localized to the hepatocytes as extrachromosomal DNA fragments. Therefore, this suggests that the bat is a potential carrier or vector of plant viruses. The present investigation on wild animals offered novel perspectives on viral invasion, variation, and host interaction.

As the only mammal that can fly, bats have organ systems with a unique morphophysiology. One of the highlights is the heart and blood circulation system, which must be able to meet the needs of blood and oxygen supply when flying. This study examined the radiography of the normal condition of the heart organ in 3 species of fruit bats, namely Cynopterus titthaecheilus, Cynopterus brachyotis and Rousettus leschenaultii using radiological silhouette analysis and clock analogy. The results showed that the heart positions of the three bat species tend to be tilted to the left with the apex moving away from the midsagittal plane. Analysis of intercostal space (ICS) value and vertebral heart score (VHS), and evaluation of radiographic features showed R. leschenaultii has a relatively larger heart size than the other two species. All three bat species have a higher VHS than mammals in general. Radiographic images obtained, and interpretation results show the position, size and normal heart parts of the three bat species. They will be useful in diagnostic efforts related to heart problems in these three species., (© 2021 Wiley-VCH GmbH.)

Uncoupling protein 1 (UCP-1) was believed to be an exclusive protein found in the brown adipose tissue of small rodents and humans; however, recent studies show that the expression of UCP-1 protein has been found in the sebaceous glands of the mouse tail and human skin. There are a few reports about the presence of UCP-1 in the sebaceous glands of other rodents, such as the Sunda porcupine (Hystrix javanica), a wild spiny rodent commonly found in Indonesia with a large sebaceous gland. The aim of this study was to identify the presence of UCP-1 in the sebaceous glands on the skin of the Sunda porcupine. The skin from three regions (thoracodorsal, lumbosacral and apex caudal) of eight adult Sunda porcupines was used to detect UCP-1-immunopositive cells through immunohistochemistry. All three regions were found immunopositive to anti-UCP-1 antibody in the sebaceous gland of quill and hair follicles, and the epidermal layer in quill and hair follicles with various intensities. The result of immunohistochemistry revealed that the thoracodorsal and apex caudal region was the most intense immunoreaction followed by the lumbosacral region. These findings proved that the presence of UCP-1 was also identified in the sebaceous glands of other rodent (Hystrix javanica) and regions of the body, which has not been reported previously.

The conventional anatomical study of specimens requires cutting processes which destruct the limited specimens. A non-destructive method, namely an ultrasonography, can be used to assess the anatomical organ information of those specimens. The aim of this research is to analyse the macroanatomy of the female reproductive organ in the Sunda porcupine (Hystrix javanica), using ultrasonographical imaging. In this study, four formaldehyde-fixed reproductive organ specimens of the Sunda porcupine were used. A 10-12 MHz linear ultrasound transducer was utilized to provide an imaging format of both longitudinal and transversal views. Photographic images were then used as comparison with a sonographic image. The results show that the ultrasound image of the Sunda porcupine reproductive organ soft tissue was hypoechoic, the lumen and antrum follicles were anechoic, while atretic follicles and the mons pubis were hyperechoic. Generally, the size of the organ was not significantly different between photographical and ultrasonographical imaging (p > .05). In conclusion, ultrasound images can be utilized for anatomical studies of the Sunda porcupine reproductive organs without destructing the specimen., (© 2020 Wiley-VCH GmbH.)

Mosquitoes transmit many kinds of arboviruses (arthropod-borne viruses), and numerous arboviral diseases have become serious problems in Indonesia. In this study, we conducted surveillance of mosquito-borne viruses at several sites in Indonesia during 2016-2018 for risk assessment of arbovirus infection and analysis of virus biodiversity in mosquito populations. We collected 10,015 mosquitoes comprising at least 11 species from 4 genera. Major collected mosquito species were Culex quinquefasciatus, Aedes albopictus, Culex tritaeniorhynchus, Aedes aegypti, and Armigeres subalbatus. The collected mosquitoes were divided into 285 pools and used for virus isolation using two mammalian cell lines, Vero and BHK-21, and one mosquito cell line, C6/36. Seventy-two pools showed clear cytopathic effects only in C6/36 cells. Using RT-PCR and next-generation sequencing approaches, these isolates were identified as insect flaviviruses (family Flaviviridae, genus Flavivirus), Banna virus (family Reoviridae, genus Seadornavirus), new permutotetravirus (designed as Bogor virus) (family Permutotetraviridae, genus Alphapermutotetravirus), and alphamesoniviruses 2 and 3 (family Mesoniviridae, genus Alphamesonivirus). We believed that this large surveillance of mosquitoes and mosquito-borne viruses provides basic information for the prevention and control of emerging and re-emerging arboviral diseases.

Wound healing in the Sunda porcupine is believed to occur quickly, although the wound is large and severe. Wound enclosure involves many processes to restore the lost or damaged skin structure where conjugated polysaccharide-protein and collagen, as the main components deposited in wound tissue to restore it. The aim of this study was to evaluate alteration of polysaccharide contents and collagen in untreated full-thickness wound healing in the thoracodorsal and lumbosacral regions in the Sunda porcupines. Histological analysis was performed by periodic acid Schiff, alcian blue pH 2.5, picrosirius red staining method and Low Vacuum Scanning Electron Microscope (LV-SEM) imaging to obtain the fundamental data of healing process. Wound healing began with re-epithelization followed by progressive wound contraction with 4 overlapping stages in about 30-50 days until the wound closed (21-30 days in thoracodorsal and 30-50 days in lumbosacral). Neutral polysaccharide was more widely distributed compared to the acid polysaccharide in almost all stages of wound healing. The ratio of collagen I to III appeared to be higher in the thoracodorsal region than the lumbosacral region during healing process. LV-SEM imaging showed changes in connective tissue structure in the wound border and granulation tissue which appeared abundant and mixed of thin and thick fiber. In conclusion, cutaneous full thickness wound healing in the Sunda porcupine occurred faster in the thoracodorsal region, which might be correlated to the role of neutral polysaccharide and a high ratio of collagen I to III.

The distribution and composition of extracellular matrix (ECM) of the spleen in two species of fruit bats, namely Cynopterus titthaecheilus and Rousettus leschenaultii, were examined by histochemistry and immunohistochemistry. Reticular fibres accompanied by laminin were identified to make up the splenic stromal network. Types I and III collagen were identified in various spleen compartments with varying intensities. Thin and short elastin fibres were scattered in several parts of the spleen. Visualization of the ECM of the spleen can better demonstrate spleen compartmentalization. The alleged vascular space structure in the fruit bats spleen was the sinus structure that was strengthened by the presence of reticular fibres that limit the sinus basement membrane. The present study identified periellipsoidal lymphoid sheath (PELS)-like structure in fruit bats spleen that had never been identified in mammals before. In addition to describing the structure, this study highlighted the variations in ECM composition of the spleen between species that can provide new insight into the phylogenetic study of spleen morphology., (© 2019 Blackwell Verlag GmbH.)

During 2016-2018, we conducted surveillance for Japanese encephalitis virus (JEV) in mosquitoes and pigs in Japan, Thailand, the Philippines, and Indonesia. Phylogenetic analyses demonstrated that our isolates (genotypes Ia, Ib, III, IV) were related to JEV isolates obtained from the same regions many years ago. Indigenous JEV strains persist in Asia.

The crude testicular cells (CTCs) contain many cell types, such as Sertoli cells, leydig cells, spermatogonial stem cells (SSCs), spermatocytes, and other somatic testicular cells, that secrete various growth factors needed in spermatogenesis. The objective of this study was to characterize development of 5-day-old mice testicular cells cultured. Crude testicular cells prepared from the testes of 5-day-old male mice were cultured in Dulbecco's Modified Eagle Medium and incubated at 37°C in a 5% CO 2 atmosphere for 6 days. The results demonstrated that the testicular cells developed rapidly with a population doubling time (PDT) of 0.63 days and more than 90% of cells were viable after being cultured for 3 days. The number of Sertoli-like cells increased significantly over days 1, 3, and 6 to 22.1%, 34.6%, and 50.1%, respectively. A significant increase was also observed in fibroblast-like cells (15.5% on day 1 to 28.8% on day 3 and to 26.6% on day 6). In contrast, the number of spermatogonia-like cells decreased significantly (54.3%, 30.4%, and 18.7%, on days 1, 3, and 6, respectively). These data indicated that the developmental pattern of the testicular cell in this study might be affected by the niche provided by the cultured testicular cells., (© 2020 Japanese Society of Animal Science.)

Ticks are one of the arthropods that play an important role in the transmission of numerous pathogens to livestock and humans. We investigated the presence of tick-borne bacteria in 23 Amblyomma varanense that fed on a water monitor (Varanus salvator) in Indonesia. Anaplasmataceae and borreliae were detected by PCR in 17.4% and 95.7% of ticks, respectively. "Candidatus Rickettsia sepangensis", spotted fever group of Rickettsia, was detected in 21.7% of ticks. The water monitor is a common reptile that is widely encountered in city areas in Asian countries. Our results suggested that Am. varanense on water monitor in Indonesia harbored several kinds of bacteria., (© 2019 The Societies and John Wiley & Sons Australia, Ltd.)

Spleen is one of the important lymphoid organs with wide variations of morphological and physiological functions according to species. Morphology and function of the spleen in bats, which are hosts to several viral strains without exhibiting clinical symptoms, remain to be fully elucidated. This study aims to examine the spleen morphology of fruit bats associated with their physiological functions. Spleen histological observations were performed in three fruit bats species: Cynopterus titthaecheilus (n = 9), Rousettus leschenaultii (n = 3) and Pteropus vampyrus (n = 3). The spleens of these fruit bats were surrounded by a thin capsule. Red pulp consisted of splenic cord and wide vascular space filled with blood. Ellipsoids in all three studied species were found numerously and adjacent to one another forming macrophages aggregates. White pulp consisted of periarteriolar lymphoid sheaths (PALS), lymphoid follicles and marginal zone. The lymphoid follicle contained a germinal centre and a tingible body macrophage that might reflect an active immune system. The marginal zone was prominent and well developed. This study reports some differences in spleen structure of fruit bats compared to other bat species previously reported and discusses possible physiological implications of the spleen based on its morphology., (© 2019 Blackwell Verlag GmbH.)

Skin becomes the largest organ in the body and protects its own inner layer. The structure and chemical composition of the skin contribute to skin condition and affect the habitat of certain bacteria. The Sunda Porcupine is one of endemic animals of Indonesia which possesses quill as the main derivate of its skin and as a defence tool against predators. The present study used nine adults (five females and four males) of Sunda Porcupine and aimed to observe the correlation of skin structure with bacterial population at the surface level. The skin was wavy due to the protrusion of quill follicle orifices on the skin surface and formed clusters. The skin of Sunda Porcupine was also wrinkled and had a lot of flakiness. Histologically, the skin was composed of epidermis, dermis, hypodermis and subcutaneous muscle. The quill follicles and their properties were the dominant structure component of the skin. No sweat gland was observed in the skin of the Sunda Porcupine, and sebaceous gland was found only around quill and hair follicles. The bacterias identified in the skin were Staphylococcus aureus, S. epidermidis, Micrococcus sp. and Salmonella sp. When compared, the bacterial population was higher in the lumbosacral region than in the thoracodorsal region, but the difference was not significant. The density of quill clusters was negatively correlated to the bacterial population. It was suggested the structure of the skin has contribution to bacterial population in dorsal trunk of the Sunda Porcupine., (© 2018 Blackwell Verlag GmbH.)

Equine piroplasmosis is an economically significant disease caused by Theileria equi and Babesia caballi, which are tick-borne hemoprotozoan parasites. Infections with these parasite species had never been reported in horses in Indonesia. The aim of the present study was to investigate the prevalence of T. equi and B. caballi in horses reared in parts of Western Java, Indonesia. Blood samples were collected randomly from 235 horses in four different districts (Bandung, Depok, Tangerang, and Bogor) in Western Java, Indonesia. Thin blood smears prepared from the sampled animals were stained by Giemsa and observed under a light microscope. Serum samples prepared from blood were screened by enzyme-linked immunosorbent assays (ELISAs) based on recombinant forms of EMA-2 and BC48 antigens to determine the seroprevalence of T. equi and B. caballi, respectively. DNA samples extracted from the same blood samples were screened by EMA-2 and BC48 gene-based nested polymerase chain reaction (nPCR) assays for T. equi and B. caballi infections, respectively. Of 235 surveyed animals, five (2.1%) and 15 (6.4%) were seropositive for T. equi and B. caballi, respectively, whereas one and four horses were nPCR-positive for T. equi and B. caballi, respectively. All of the surveyed animals were negative for T. equi and B. caballi by microscopy. The T. equi EMA-2 and B. caballi BC48 gene fragments amplified by the nPCR assays were cloned, sequenced, and subjected to bioinformatic and phylogenetic analyses. The T. equi EMA-2 gene sequence from an Indonesian horse was identical to sequences from Florida and Washington strains and clustered together with these sequences in phylogeny. On the other hand, four Indonesian BC48 gene sequences shared 99.8-100% identity scores. This present study is the first to report T. equi and B. caballi in horses in Indonesia. Our findings highlight the need for monitoring horses in Indonesia for clinical piroplasmosis caused by T. equi and B. caballi., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Flying foxes belonging to the genus Pteropus are known to be reservoirs of zoonotic viruses. In this study, we describe the isolation of Pteropine orthoreovirus (PRV) from rectal swab samples of Pteropus vampyrus in Indonesia. PRV is an emerging zoonotic respiratory virus that can be transmitted from bats to humans. Rectal swabs (n = 91) were screened by PCR for PRV and 10 (11%) were positive. Phylogenetic analysis based on nucleotide sequences indicated that the S2, S3, S4, M3, L2, and L3 segments of one isolate (Garut-69) were closely related to previously isolated strains in Indonesia. The remaining gene segments showed both similarity and genetic divergence with other PRV strains, suggesting that re-assortment events had occurred. This is the first report of PRV infection to P. vampyrus in West Java, Indonesia.

Flying foxes are important in the maintenance of forests and diversity. However, knowledge of their behavioral ecology, especially of movement and foraging patterns, which are essential for conservation and management of their populations, are not well known. Therefore, movement behavior of two individuals of Pteropus vampyrus were examined using an Argos telemetry system, and foraging pattern of Pteropus spp. was directly observed, at West Java province, Indonesia in October 2017. The maximum distance between the location at which bats were released and their furthest roost, recorded via satellite telemetry, was approximately 100 km. This reflects the long-distance flight ability of P. vampyrus. Daytime roosting sites and nighttime foraging places consisted of several types of habitats, such as intact forests, agricultural lands, and residential areas. This evidence indicated that there was habitat overlap between humans and bats in West Java province. According to direct observation of the behaviors of flying foxes at two locations within residential areas, various activities such as wing spreading, excretion, fighting, aggressive calls, movement, hanging relax, and hanging alert were found. The number of bat-visits to the trees varied among night hours, and had a positive correlation with the number of fruit dropping. The data obtained in this study have improved our understanding of nighttime behavior and habitat utilization of P. vampyrus, that can be used to support landscape management, species conservation, and disease prevention in regions of Southeast Asia.

Stages of the seminiferous epithelium of the testis of the wild Javan muntjac ( Muntiacus muntjak muntjak ) in hard antler period were characterized based on the tubular morphology method. The number and the relative frequencies of seminiferous epithelium stages and the morphometry of germinal cell nuclei were identified microscopically. We identified eight stages of seminiferous epithelium in testicular tissue of the Javan muntjac and found that the relative frequencies of stages I to VIII were 14.87, 15.12, 17.75, 6.87, 7.37, 12.37, 13, and 12.62%, respectively. The diameter of the nuclei of germinal cells varied in each stage of seminiferous epithelium. Diplotene-stage primary spermatocytes had prominent and large nuclei ~8.97 ± 1.0 μ m in stages III and IV. Pachytene primary spermatocytes appeared in most stages, except stage IV, whereas leptotene- and diplotene-stage primary spermatocytes were found in stages I and II, and III and IV, respectively. Round spermatids were observed in stages IV to VIII and in stage I but were absent in stages II and III, while elongated spermatids were observed in all stages except stage I. Our findings show that the stages of seminiferous epithelium in the Javan muntjac are similar to those found in neotropical cervids, small ruminants, and other domestic animals.

Flying foxes have been considered to be involved in the transmission of serious infectious diseases to humans. Using questionnaires, we aimed to determine the direct and/or indirect contacts of flying foxes in an Indonesian nature conservation area with domestic animals and humans living in the surrounding area. We surveyed 150 residents of 10 villages in West Java. Villages were classified into 3 groups: inside and/or within 1 km from the outer border of the conservation area and 1-5 km or 5-10 km away from the reserve's outer border. Data were collected by direct interview using a structured questionnaire consisting of the respondent characteristics (age, sex and occupation); histories of contacts between flying foxes and humans, dogs and other domestic animals; and knowledge about infectious diseases, mainly rabies, in flying foxes. We found that flying foxes from the nature conservation area often enter residential areas at night to look for food, especially during the fruit season. In these residential areas, flying foxes had direct contacts with humans and a few contacts with domestic animals, especially dogs. People who encounter flying foxes seldom used personal protective equipment, such as leather gloves, goggles and caps. The residents living around the conservation area mostly had poor knowledge about flying foxes and disease transmission. This situation shows that the population in this region is at a quite high risk for contracting infectious diseases from flying foxes.

Flying foxes, the genus Pteropus, are considered viral reservoirs. Their colonial nature and long flight capability enhance their ability to spread viruses quickly. To understand how the viral transmission occurs between flying foxes and other animals, we investigated daytime behavior of the large flying fox (Pteropus vampyrus) in the Leuweung Sancang conservation area, Indonesia, by using instantaneous scan sampling and all-occurrence focal sampling. The data were obtained from 0700 to 1700 hr, during May 11-25, 2016. Almost half of the flying foxes (46.9 ± 10.6% of all recorded bats) were awake and showed various levels of activity during daytime. The potential behaviors driving disease transmission, such as self-grooming, mating/courtship and aggression, peaked in the early morning. Males were more active and spent more time on sexual activities than females. There was no significant difference in time spent for negative social behaviors between sexes. Positive social behaviors, especially maternal cares, were performed only by females. Sexual activities and negative/positive social behaviors enable fluid exchange between bats and thus facilitate intraspecies transmission. Conflicts for living space between the flying foxes and the ebony leaf monkey (Trachypithecus auratus) were observed, and this caused daily roosting shifts of flying foxes. The ecological interactions between bats and other wildlife increase the risk of interspecies infection. This study provides the details of the flying fox's behavior and its interaction with other wildlife in South-East Asia that may help explain how pathogen spillover occurs in the wild.

We successfully established a mass production system for an influenza virus-like particle (VLP) vaccine using a synthetic H5 hemagglutinin (HA) gene codon-optimized for the silkworm. A recombinant baculovirus containing the synthetic gene was inoculated into silkworm pupae. Four days after inoculation, the hemagglutination titer in homogenates from infected pupae reached a mean value of 0.8 million hemagglutination units (HAU), approximately 2,000 μg HA protein per pupa, more than 50-fold higher than that produced with an embryonated chicken egg. VLPs ranging from 30 nm to 300 nm in diameter and covered with a large number of spikes were detected in the homogenates. The spikes were approximately 14 nm long, similar to an authentic influenza HA spike. Detailed electron micrographs indicated that the VLP spike density was similar to that of authentic influenza virus particles. The results clearly show that the expression of a single HA gene can efficiently produce VLPs in silkworm pupae. When chickens were immunized with the pupae homogenate, the hemagglutination inhibition titer in their sera reached values of 2,048-8,192 after approximately 1 month. This is the first report demonstrating that a large amount of VLP vaccine could be produced by single synthetic HA gene in silkworm pupae. Our system might be useful for future vaccine development against other viral diseases., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The morphology of the stomach of Malayan pangolin, Manis javanica was studied at macroscopic, light microscopic, and scanning electron microscopic levels. The stomach of M. javanica was C-shaped with short lesser curvature. At the oesophageal junction, the inner smooth muscle was thickened in the greater curvature side. The entire stomach was lined by a thick cornified stratified squamous epithelium, except at the duct orifices of glands and in the pyloric gland region. The wall of the fundus was thin and devoid of glands. The gastric glands consisted of mucous, oxyntic, and pyloric glands. The mucous glands were observed in the lesser curvature (Mg-L), in the greater curvature (Mg-G), and in the pyloric canal (Mg-C) respectively. The oxyntic glands were organized into gland mass, making an oval mound elevated to the gastric lumen, in the middle of the greater curvature. The oxyntic gland mass has a single common duct with opening directed to the pyloric side. This duct was surrounded by mucus gland (Mg-G). The pyloric glands were located caudal to the pylorus. There was no sphincter at the pyloric-duodenal junction. Large mucosal protuberance, the torus pyloricus was observed in the side of the lesser curvature of the pyloric canal. In the lumen of pyloric canal region, numerous spines and small pebbles were observed. The muscle layers in the wall of this region were considerably thickened. The present results on the stomach of M. javanica are thought to be closely related to the toothless and eating habits of this animal species., (© 2010 Blackwell Verlag GmbH.)

The dorsal lingual surface of the lesser dog-faced fruit bat was examined by scanning electron microscopy (SEM). Filiform (Fi), fungiform (Fu) and vallate papillae (V) were observed. The Fi papillae were distributed over the entire dorsal surface of the tongue. The Fi papillae notably differed in morphology by their location on the tongue and could be classified into 5 types: 1) crown-like papillae, 2) giant trifid papillae, 3) scale-like papillae, 4) small conical papillae, and 5) large conical papillae. The Fu papillae were present rounded bodies on the anterior 2/3 of the tongue. The lesser dog-faced fruit bat showed the triangular arrangement of the three V, with the apex of the triangle directed posteriorly.

The cycle of the seminiferous epithelium in the Java fruit bat, Pteropus vampyrus, and the Japanese lesser horseshoe bat, Rhinolophus cornutus, was investigated by light microscopy and the characteristics of spermiogenesis were compared between these two species. In the Java fruit bat, the cycle of the seminiferous epithelium was divided into 11 stages and developing spermatids were subdivided into 13 steps. While in the Japanese lesser horseshoe bat, the cycle of the seminiferous epithelium was divided into 10 stages and developing spermatids were subdivided into 13 steps. Excepting slight morphological differences, the characteristics of acrosomal formation in both species were almost similar with each other. In the Java fruit bat after stage VII, the acrosome gradually elongated, flattened and finally became scoop-like in shape. In the Japanese lesser horseshoe bat after stage VIII, the acrosome elongated, flattened and then slightly shortened. Before spermiation, the acrosome became long spatula-like in shape. The elongation and flattening of spermatids in these two species were similar to those in insectivores. The finding may reflect the fact that the order Chiroptera is phylogenetically close to the order Insectivora.

Lectin-binding patterns in the testes of the northern smooth-tailed tree shrew, Dendrogale murina and Java tree shrew, Tupaia javanica were studied by light microscopy and compared the data with those of the common tree shrew. Four lectins (PNA, SBA, BPA and GS-II) were used in this study. Peanut (Arachis hypogaea) agglutinin (PNA), soybean (Glycine max) agglutinin (SBA) and Bauhinia purpurea agglutinin (BPA) showed a strong reaction in the acrosomal region from Golgi to acrosome-phase spermatids in three species of tree shrews. These lectins also showed a granular positive reaction in the cytoplasm from acrosome to maturation-phase spermatids in three species, except that BPA revealed no granular reaction (though it was positive) in the spermatid cytoplasm of the northern smooth-tailed tree shrew and that PNA revealed no reaction in the spermatid cytoplasm of the common tree shrew. While, Griffonia simplicifolia-II agglutinin (GS-II) showed a positive reaction in the acrosomal region of Golgi-phase spermatids in three species of tree shrews. Although GS-II was positive in the spermatocyte cytoplasm of three species, it showed granular in the northern smooth-tailed tree shrew and common tree shrew but not granular in the Java tree shrew. Thus, the lectin-binding patterns in testes were similar among three species belonging to the Order Scandentia. However, slight differences were also detected even among these phylogenetically-close species.

The lesser mouse deer (Tragulus javanicus) is one of the most primitive ruminants. Skulls of lesser mouse deer were measured to evaluate their growth. The age was estimated from the eruption and attrition of molars on the mandible. A total of 95 specimens was divided into five age groups, and the growth pattern in each sex was established for some measurements. The relative growth coefficients were calculated for the head and body length and several parts of the cranium and mandible against the profile length. Sexual dimorphism could not be found on the skull in any age group except for measurements related to the upper canine. However, females showed a higher value in head and body length in the oldest group. This feature was consistent with an interpretation of mouse deer society as monogamous. Growth was more rapid in length than in width. Various statistical analyses showed that the visceral cranium related to masticatory facility was much better developed than the neurocranium. However, observations on the shape of the mandible ramus and of the premolars implied that the masticatory function was not suitable for taking roughage. This speculation agreed with previous reports on the feeding habits and digestive organs of the mouse deer.

The dorsal lingual papillae of the lesser mouse deer were studied morphologically using light and scanning electron microscopy. Four types of papillae, filiform, fungiform, vallate and foliate, were observed. Filiform papillae consisted of larger main papillae with smaller secondary papillae on their anterolateral aspects. Secondary papillae were well distributed over the anterior two-thirds of the tongue, but were very rare or absent in the posterior third. Fungiform papillae were distributed among the filiform papillae, being larger and more abundant on the tip of the tongue. Vallate papillae were round-flat or long-flat, surrounded by a prominent circular groove and a thin annular pad. An important finding was the presence of distinct and prominent foliate papillae on the posterolateral sides of the tongue. Keratinisation of the covering stratified squamous epithelium was relatively weak. Taste buds were observed in the epithelium of the fungiform, vallate and foliate papillae. The lingual papillae of the lesser mouse deer showed some characteristics that differed from those reported for domestic ruminants. These may be related to the feeding habits and the type of food eaten by this species.

The stomach of the lesser mouse deer was studied at the light microscopic level using histological and immunohistochemical methods. The stomach was clearly differentiated into rumen, reticulum including reticular groove, a small transition zone and abomasum. The mucosal surface of the rumen, reticulum and transition zone was lined with a stratified squamous epithelium and that of the abomasum with a simple columnar type. The epithelial keratinization was weak in the rumen, floor of the reticular groove and transition zone, while it was strong in the reticulum, especially on the tip of the reticulum papillae. Large sinusoidal capillaries were often present in the ruminal papillae. In the ruminal mucosa, a thin layer of alpha-smooth muscle actin immunoreactive cells was demonstrated by immunohistochemistry. The muscularis mucosae of the reticulum was continuous and well-developed. The transition zone appeared as a nonglandular area having many low mucosal folds and two layers of tunica muscularis. The abomasal mucosa consisted of cardiac, proper gastric and pyloric glands. Cells immunoreactive for bovine pepsinogen and bovine prochymosin antisera were demonstrated in the abomasum. It is suggested that the characteristic features observed might be adaptations to a relatively rapid passage and rapid absorption of the fermentation products. There is some evidence that the transition zone is not a part of either the floor of the reticular groove or the abomasum, suggesting a possible reevaluation of the term used for the reticulo-abomasal orifice in the mouse deer.