Carnivores (Mammalia: Carnivora) in South China: a status review with notes on the commercial trade

• 1 Evidence from the literature, interviews, market surveys, general rapid field surveys and camera trapping was reviewed to infer the regional status of 33 recorded carnivores in South China (Guangxi, Guangdong, Hainan, Hong Kong and Macau).
• 2 The carnivore fauna in South China is among the most depleted for any continental area in the world.
• 3 The tiger Panthera tigris, leopard Panthera pardus, grey wolf Canis lupus and binturong Arctictis binturong are probably extirpated.
• 4 The dhole Cuon alpinus, Asiatic golden cat Catopuma temmincki and clouded leopard Neofelis nebulosa are at great risk of regional extirpation, as are the more ecologically adaptable red fox Vulpes vulpes, raccoon dog Nyctereutes procyonoides, Asian badger Meles leucurus, Eurasian otter Lutra lutra, Oriental small‐clawed otter Aonyx cinereus and large Indian civet Viverra zibetha.
• 5 Though more widely reported, the Asiatic black bear Ursus thibetanus, yellow‐throated marten Martes flavigula, hog badger Arctonyx collaris and Siberian weasel Mustela sibirica are at regional risk.
• 6 The status of the Burmese ferret‐badger Melogale personata, stripe‐backed weasel Mustela strigidorsa, smooth‐coated otter Lutrogale perspicillata, large‐spotted civet Viverra megaspila and Owston's palm civet Chrotogale owstoni is uncertain due to inadequate information.
• 7 Five carnivore species – the Chinese ferret‐badger Melogale moschata, masked palm civet Paguma larvata, yellow‐bellied weasel Mustela kathiah, small Indian civet Viverricula indica and leopard cat Prionailurus bengalensis– are relatively secure (though not common) in the region, along with the Asian palm civet Paradoxurus hermaphroditus and the Javan mongoose Herpestes javanicus in some areas, while the spotted linsang Prionodon pardicolor and crab‐eating mongoose Herpestes urva may have been under‐reported.
• 8 The main threats have been habitat destruction and unsustainable exploitation. Only in Hong Kong, where enforcement of protection legislation has been stronger, are the surviving carnivore species easily encountered.
• 9 Improved enforcement and monitoring are essential to retaining the remainder of South China's carnivores. Support could be boosted by conservation education and better research.

     

Interchange as the main factor determining wildlife–vehicle collision hotspots on the fenced highways: spatial analysis and applications

Despite the fact that all highways in Hungary are built with fencing, there are still 5 % of traffic accidents that involve wildlife. Therefore, this study focused on the incidence of collisions along fenced roads. Wildlife–vehicle collision (WVC) hotspots were mapped, and the spatial frequencies were analysed with Poisson regression. In general, most WVCs and almost all of the roe deer fatalities occurred at highway intersections, or at interchanges. Red fox casualties also occurred at interchanges as well as at passages. Wild boar fatalities were not particularly frequent at interchanges but were recorded near railways that are parallel to highways; otter–vehicle collision hotspots were found near their habitats and migration corridors such as streams. For otter, badger and wild boar, we were able to examine the role of local population density; most WVCs happened in areas of high population density. The badger model predicted that badger kills were more likely where the fence was not buried in the soil. Most WVCs occur at interchanges because wildlife enters the right-of-way (ROW) at fence ends; or it enters at a fence gap and runs along the outside of fence and becomes funnelled onto the ROW at the interchanges. Interruption in the continuity and linearity are important factors in both cases. We concluded that the number of WVCs can be reduced significantly if animals were prevented from entering highway interchanges and proposals for mitigation were made. We also propose a tool to assist in alleviating WVCs, by mapping them in Google Maps and integrating hotspots into a car navigation system.     

Prevalence of embedded shotgun pellets in protected and in legally hunted medium-sized carnivores in Denmark

Illegal killings of protected species may significantly affect their population status or lower recovery rates after protection. Poaching is difficult to monitor directly, hence indirect methods are required to document poaching incidences and levels. The use of shotguns inevitably results in wounding some animals that are not killed. Inefficient hunting methods may result in high annual infliction rates in game species. To reduce wounding of game species, an action plan was implemented in Denmark. Using X-ray analysis, we monitor trends in prevalence of embedded shotgun pellets in the legally hunted red fox (Vulpes vulpes) after the implementation of the action plan, and assess poaching levels of two protected species, the otter (Lutra lutra) and the badger (Meles meles) in Denmark. Overall, 13.8% of the red foxes and 6.9% of the otters carried embedded shotgun pellets, while none were recorded in badgers. The prevalence of embedded shotgun pellets in red foxes declined from 24.9% to 8.5% from the late 1990s to the late 2000s, while no change was recorded in otters. Prevalence of embedded shotgun pellets was similar in otters and red foxes collected in the late 2000s. The decline in wounded red foxes suggests that the hunting methods have changed after the action plan was implemented in Denmark. The high proportion of otters carrying embedded shotgun pellets shows that there is a significant poaching of this protected species in Denmark, possibly by hunters and fish farmers that cannot distinguish between otter and non-native American mink (Neovison vison).     

Antibodies to Mycobacterium bovis in wild carnivores from Doñana National Park (Spain)

We conducted a retrospective serologic survey for antibodies against the MPB70 protein of Mycobacterium bovis in wild carnivores from Do?ana National Park (southwestern Spain). Serum samples from 118 red foxes (Vulpes vulpes), 39 Iberian lynx (Lynx pardinus), 31 Eurasian badgers (Meles meles), five Egyptian mongoose (Herpestes ichneumon), four European genet (Genetta genetta), and one Eurasian otter (Lutra lutra) were analyzed using an indirect competitive enzyme-linked immunoassay. Antibodies against the MPB70 protein of M. bovis were detected in seven badgers, five foxes, and one lynx. The frequency of positive animals was significantly higher in badger (23%) than in lynx (3%) and fox (4%). Antibodies were not detected in other species. Annual antibody frequency peaked at 38% in badgers and 11% for red fox. These species may contribute to persistence of bovine tuberculosis in Do?ana.     

Carnivore population trends in Spanish agrosystems after the reduction in food availability due to rabbit decline by rabbit haemorrhagic disease and improved waste management

We conducted spotlight counts from 1992 to 2006 in 59 localities to describe carnivore presence, distribution and relative abundance in open agriculture lands in Aragon, northeastern Spain. During the study period, urban waste and carcases of domestic livestock became less available to wild animals, and rabbit (Oryctolagus cuniculus) abundance was low after rabbit haemorragic disease. We calculated a kilometric abundance index (KAI, individuals seen per 100 km surveyed) and the ratio between the number of positive localities in which a species was detected and the total of localities surveyed in a year. Indices to abundance and presence included: red fox (Vulpes vulpes), KAI per 100 km 10.70 and 100.0% localities per year; stray dog (Canis familiaris), KAI 0.48 and 22.6% localities; wildcat (Felis silvestris), KAI 0.10 and 19.1% localities; domestic cat (Felis catus), KAI 0.21 and 20.6% localities; polecat (Mustela putorius), KAI 0.02 and 2.0% localities; badger (Meles meles), KAI 0.16 and 23.6% localities; stone marten (Martes foina), KAI 0.11 and 19.6% localities; weasel (Mustela nivalis), KAI 0.002 and 0.6% localities; and genet (Genetta genetta), KAI 0.06 and 10.0% localities. Indices of badger abundance increased significantly, whereas indices of stray dogs decreased significantly over the study period. Abundance indices of red fox, the most important predator of small game, were stable throughout the study period. No significant trend was observed to other studied species. This study concludes that Aragon region maintains a diverse and rather stable carnivore community and shows that large scale data may help to identify trends of the more abundant wild and feral carnivores.     

Burrowing by badgers (Meles meles) and foxes (Vulpes vulpes) changes soil conditions and vegetation in a European temperate forest

This study examined the effects of burrow digging and habitation by the European badger (Meles meles) and the red fox (Vulpes vulpes) on soil properties and the plant community. The vegetation of control plots located in a similar but undisturbed habitat was compared with that of 18 burrow plots established at badger setts (N = 9) and fox dens (N = 9) in a lowland forest area in Poland. Soil physicochemical properties at different disturbance levels (mounds, intermounds and reference areas) were also investigated. The animals altered nutrient availability in the burrow plots considerably by excavating material from deep soil horizons that were less acidic and higher in K, Ca, Mg and available P but poorer in C and N. The effect was stronger for the badger, probably because it displaced larger amounts of material and disturbed wider areas. The activity of the two carnivores induced similar changes in plant communities. They increased herbaceous species richness and caused a shift in the herbaceous species composition: versus the control plots, the burrow plots contained more fugitive species (short-living plants typical for disturbed environments), among which ruderal forbs, including nutrient-demanding species, dominated. The carnivores also increased the species richness of fleshy-fruited shrubs and trees. The primary reason for this was probably not burrowing but endozoochorous seed dispersal. Overall, the results indicate that the badger setts and fox dens differ significantly from the forest matrix in terms of soil and vegetation parameters, and that they contribute to habitat heterogeneity and biological diversity.     

The diet of feral raccoon dog (<Emphasis Type="Italic">Nyctereutes procyonoides</Emphasis>) and native badger (<Emphasis Type="Italic">Meles meles</Emphasis>) and red fox (<Emphasis Type="Italic">Vulpes vulpes</Emphasis>) in Denmark

The raccoon dog (Nyctereutes procyonoides) is an East Asian Canid that has been introduced in Europe. Introduction of alien species is an increasing conservation issue. We examined the diet of a recently established raccoon dog population in Denmark by analysing stomach content in 249 carcasses collected in 2008–2016. Raccoon dog diet was compared to the diet of native badger (Meles meles) and red fox (Vulpes vulpes) in Denmark. The most common food for raccoon dogs were invertebrates (frequency of occurrence, FO 69%), small mammals (FO 68%), birds (FO 41%), fruits (FO 38%), amphibians (FO 36%) and carrions (FO 34%). The occurrence of invertebrates was highest during spring and summer, while fruits, cereals and carrions were eaten most often during autumn and winter. As expected, raccoon dog shared the major food categories with badger and red fox, but generally, it had a wider dietary niche. Overall, dietary overlap between raccoon dog and badger was 0.74 (Pianka index, O_jk). The dietary overlap with red fox was relatively high in all seasons, peaking in summer (O_jk 0.87) and dropping in winter (O_jk 0.79). Despite the dietary overlap between the alien racoon dog and native red fox and badger, the species may coexist due to partitioning of feeding habitats and/or because the red fox is limited by other factors, e.g. diseases and anthropogenic activities. The introduced raccoon dog seems to fit a dietary niche between badger and red foxes in human-dominated landscapes in north-western Europe.     

Spatial and temporal adjustments allowing the coexistence among carnivores in Liguria (N-W Italy)

We investigated the seasonal spatial and temporal co-occurrence of three carnivore species in Liguria region (NW Italy)—the red fox (Vulpes vulpes), the European badger (Meles meles) and the wolf (Canis lupus)—using the information provided by camera-trapping monitoring. Data were collected from January 2013 to January 2015 by positioning camera traps in 200 sample stations. During 3479 trap days, we collected 1048 independent videos of target carnivore species, which revealed a general spatial coexistence among carnivores with some differences in seasonal occurrence of species. The red fox and the European badger showed temporal segregation, as their activity patterns suggested a differential use of night-time in all seasons. Activity patterns of the red fox and the wolf revealed moderate-high overlap and similar density distributions in all seasons except during winter. Coexistence between these species may be allowed by temporal segregation during winter and spatial segregation during spring. Finally, results regarding the European badger and the wolf suggest a moderate temporal segregation with a marked avoidance effect for the European badger induced by the presence of tracks left by wolves. Programmes aimed at carnivore conservation, and management should treat the entire guild, as it has been demonstrated that populations of different carnivores interact with each other in complex ways and that fine-scale mechanisms regulating carnivore assemblage influence different aspects of natural communities.     

Molecular Characterization of Enterocytozoon bieneusi in Wild Carnivores in Spain

Microsporidia comprises a diverse group of obligate intracellular parasites that infect a broad range of invertebrates and vertebrates. Among Microsporidia, Enterocytozoon bieneusi is the most frequently detected species in humans and animals worldwide bringing into question the possible role of animal reservoirs in the epidemiology of this pathogen. Although E. bieneusi is an emerging zoonotic pathogen able to infect many domestic and wild mammals that could act as reservoir of infection for humans and other animals, only few studies have documented its occurrence in wild carnivores. To determine the occurrence of E. bieneusi in wild carnivores, we examined 190 wild carnivores collected from different locations in Spain. Twenty‐five fecal samples (13.2%) from three host species (European badger, beech marten, and red fox) were E. bieneusi‐positive by PCR. Nucleotide sequence analysis of the ITS region revealed a high degree of genetic diversity with a total of eight distinct genotypes including four known (PtEbIX, S5, S9, and WildBoar3) and four novel (EbCar1‐EbCar4) genotypes identified. Phylogenetic analysis showed that the four novel genotypes (EbCar1‐EbCar4), S5, S9, and WildBoar3 clustered within the previously designated zoonotic Group 1. Our results demonstrate that human‐pathogenic genotypes are present in wild carnivores, corroborating their potential role as a source of human infection and environmental contamination.     

Fruit consumption by carnivores in Mediterranean Europe

• 1 We reviewed patterns of fruit consumption amongst 10 species of mesocarnivores: red fox Vulpes vulpes, weasel Mustela nivalis, stoat Mustela erminea, polecat Mustela putorius, stone marten Martes foina, pine marten Martes martes, Eurasian badger Meles meles, common genet Genetta genetta, Egyptian mongoose Herpestes ichneumon and wildcat Felis silvestris in Mediterranean Europe.
• 2 The 65 reviewed studies recorded 79 different fruits eaten by carnivores, 58 of which were identified to species. Most records (63%) were of fleshy fruits with high pulp content. The frequency of occurrence of fruit items varied widely amongst species and regions. Four of the carnivore species (red fox, stone marten, badger and common genet) included more than 30 fruit species in their diet.
• 3 A longitudinal pattern was detected in the consumption of fruit in the Mediterranean region, with the frequency of occurrence of fruit consumption increasing towards the east.

     

Theobromine intoxication in a red fox and a European badger in Sweden

A red fox (Vulpes vulpes) and a European badger (Meles meles) were found dead on a golf-course in October 1997 near Stockholm (Sweden). At necropsy, both animals were obese and the main finding was acute circulatory collapse. Theobromine intoxication was suspected as chocolate waste was available at a nearby farm and no other cause of death could be detected. Gastric contents and samples of liver from both animals were analyzed by reversed-phase high pressure liquid chromatography for the presence of methylxanthines. Theobromine and caffeine were detected in gastric contents and theobromine was identified in the liver samples from both animals. This appears to be the first report of theobromine intoxication in the red fox and the European badger.     

Evaluation of scent-station surveys to monitor the distribution of three European carnivore species (Martes foina, Meles meles, Vulpes vulpes) in a fragmented landscape

Scent-stations are frequently used in North and South America to monitor carnivore populations, and their efficacy has been widely tested on various species. On the other hand, scent-stations have rarely been used to monitor European species and their efficacy has never been tested. Therefore, an acritical use on Eurasiatic species may lead to biased results, but also a neglection of this technique may limit methodological improvements.We evaluated scent-stations detection efficacy on three eurasiatic species: badger (Meles meles), fox (Vulpes vulpes) and beech marten (Martes foina) using data from a carnivore patch-use survey conducted in a fragmented landscape in the Province of Siena, central Italy. Scent-station survey data was compared with distribution data obtained from the utilisation of different techniques (track and camera trap surveys and interviews with local people) and an estimate of proportion of sites occupied inferred through the application of occupancy models.We show how by using scent-stations, fox distribution was largely underestimated, badger distribution was biased (this species was missed in small fragments) while beech marten (Martes foina) distribution was relatively unbiased. We conclude that application of this technique should follow a pilot study and cost/benefits should be carefully considered.     

Mercury in wild terrestrial carnivorous mammals from north-western Poland and unusual fish diet of red fox

Total mercury concentrations were determined in the kidney (K), liver (L), and pectoral muscle (M) of 19 individuals representing wild carnivorous mammals from NW Poland: 10 red foxes Vulpes vulpes (Linnaeus, 1758), 3 raccoon dogs Nyctereutes procyonoides Gray, 1834, 2 badgers Meles meles Linnaeus, 1758, 3 pine martens Martes martes Linnaeus, 1758, and 1 polecat Mustela putorius Linnaeus, 1758. The sample of red fox included 3 immature specimens found on Mielin Island; the island supports a black cormorant colony, and the foxes found there had fed mostly on cormorant nestlings as well as on fish and their remains. In addition to the Mielin Island foxes, the group of foxes included 3 other immature and 4 adult individuals. The highest mean of mercury concentrations were revealed in the Mielin red fox juveniles: 5.11, 4.52, and 1.56 mg/kg d.w. being recorded in K, L, and M. No significant differences in mercury concentrations in the respective tissues were found between the remaining immature and adult red foxes; their mercury concentrations were several times lower than those of the Mielin individuals. In all the animals except the Mielin foxes, mercury concentrations in K, L, and M did not exceed 1.3, 1.0 and 0.5 mg/kg d.w., respectively, the highest values being in badgers (which feed mostly on soil invertebrates), followed by pine martens and then the canids (red fox and raccoon dog). Studies on common and widely distributed terrestrial animals, particularly red fox and badger, may provide numerous valuable comparative data on mercury contamination of different areas of the northern hemisphere.     

Adiaspiromycosis in Czechoslovakian mammals

The examination of 46 mammalian species in Czechoslovakia revealed adiaspiromycosis caused by the Emmonsia crescens in 22 species of free living animals; E. parva was the etiological agent in 3 species. The disease was most frequently found in carnivores of the family Mustelidae.     

Estimates of regional population densities of badger Meles meles, fox Vulpes vulpes and hare Lepus europaeus using walked distance sampling

Walked spotlight transect surveys with distance sampling were used to estimate regional population densities of badger (Meles meles), fox (Vulpes vulpes) and brown hare (Lepus europaeus) in south-west England (Cornwall, Devon, Gloucestershire, Herefordshire) and Wales (Pembrokeshire, Borders, North Wales). All regions were surveyed during spring 2006 with English regions re-surveyed in autumn 2006. In each region, surveys were conducted in a random sample of 19.6 km² areas (mean areas per region: spring = 19, autumn = 25). Within each survey area, a semi-random transect was established in each of a random sample of fields (open habitat almost exclusively pasture). Transects were subsequently walked at night with spotlights (mean transects per survey area: spring = 21, autumn = 21). Each area was surveyed twice during a season. Total transect length per region ranged from 137 to 193 km in spring and 230 to 250 km in autumn. The mean density of species per region was: badger 1.5–4.8 km⁻², fox 1.0–4.0 km⁻², hare 0.4–4.6 km⁻². The study has provided baseline estimates of regional densities against which any future equivalent surveys can be compared. It has also illustrated the practical application of large-scale walked distance sampling to surveys of British mammals.     

Two’s company,three’s a crowd: Exploring how host–parasite–microbiota interactions may influence disease susceptibility and conservation of wildlife

A large body of research has demonstrated that host‐associated microbiota—the archaeal, bacterial, fungal and viral communities residing on and inside organisms—are critical to host health (Cho & Blaser, 2012). Although the vast majority of these studies focus on humans or model organisms in laboratory settings (Pascoe, Hauffe, Marchesi, & Perkins, 2017), they nevertheless provide important conceptual evidence that the disruption of host‐associated microbial communities (termed “dysbiosis”) among wild animals may reduce host fitness and survival under natural environmental conditions. Among the myriad of environmental factors capable of inducing dysbiosis among wild animals (Trevelline, Fontaine, Hartup, & Kohl, 2019), parasitic infections represent a potentially potent, yet poorly understood, factor influencing microbial community dynamics and animal health. The study by DeCandia et al. in this issue of Molecular Ecology is a rare example of a host–parasite–microbiota interaction that impacts the health, survival and conservation of a threatened wild animal in its natural habitat. Using culture‐independent techniques, DeCandia et al. found that the presence of an ectoparasitic mite (Otodectes cynotis) in the ear canal of the Santa Catalina Island fox (Urocyon littoralis catalinae) was associated with significantly reduced ear canal microbial diversity, with the opportunistic pathogen Staphylococcus pseudintermedius dominating the community. These findings suggest that parasite‐induced inflammation may contribute to the formation of ceruminous gland tumours in this subspecies of Channel Island fox. As a rare example of a host–parasite–microbiota interaction that may mediate a lethal disease in a population of threatened animals, their study provides an excellent example of how aspects of disease ecology can be integrated into studies of host‐associated microbiota to advance conservation science and practice.     

Temporal and geographical variation in skull size of the red fox (Vulpes vulpes) and the Eurasian badger (Meles meles) in Austria

Using museum material, we studied temporal changes in skull size of 185 red foxes (Vulpes vulpes Linnaeus, 1758) and 71 Eurasian badgers (Meles meles Linnaeus, 1758) collected in Austria between 1866 and 2007. Four measurements were taken and combined into principal components by means of a principal components analysis. Akaike's information criterion models indicated that skull size of the red fox and the Eurasian badger is positively related to the year of collection and negatively to altitude but not to latitude or longitude. However, for the Eurasian badger, the relationship between skull size and year is only on the verge of significance. We suggest that the increase in skull size with year is a result of improved food availability during the 20th Century from man‐made resources such as agricultural produce and garbage, and the decrease in skull size with altitude is possibly the result of a parallel decrease in primary productivity. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, ●● , ●●–●●.     

National predictors of hedgehog Erinaceus europaeus distribution and decline in Britain

We utilise a volunteer survey recording roadkills between 2001 and 2011 to examine the factors affecting hedgehog Erinaceus europaeus abundance and decline. Hedgehogs were most abundant in the North and East of England and in Scotland, regions characterised by low badger numbers. Hedgehogs selected arable land and urban areas relative to their availability. Badger Meles meles and fox Vulpes vulpes abundance were negatively associated with hedgehog abundance at the 10 km² scale. At the county level, foxes were positively associated with hedgehog numbers and badgers negatively associated. The mechanism behind the relationships between hedgehogs and badgers and foxes merits further investigation.     

Molecular detection of coccidian Apicomplexa Parasites isolated from wild crab-eating and pampas foxes through novel TaqMan™ probes: a contribution to their molecular epidemiology

Neospora caninum, Toxoplasma gondii and Hammondia spp. are coccidian parasites similar in morphology. Molecular techniques are necessary to detect parasite DNA isolated from stool samples in wild canids because they were reported as definitive hosts of N. caninum life cycle. The objective of this study was to develop a highly sensitive and accurate molecular method for the identification of coccidian Apicomplexa parasites in crab-eating fox (Cerdocyon thous) and pampas fox (Lycalopex gymnocercus). Tissue samples from road-killed animals (pampas fox?=?46, crab-eating fox?=?55) and feces (pampas fox?=?84, crab-eating fox?=?2) were collected, and species were diagnosed through molecular assay. PCR was used for the amplification of a fragment of the coccidian Apicomplexa nss-rRNA gene. Additionally, we developed a novel real-time PCR TaqMan? probe approach to detect T. gondii- Hammondia spp. and N. caninum. This is the first report of N. caninum DNA in pampas fox feces (n?=?1), thus it was also detected from pampas fox tissues (n?=?1). Meanwhile, T. gondii was found in tissues of pampas (n?=?1) and crab-eating (n?=?1) foxes and H. triffittae in one crab-eating fox tissue. Despite the low percentage (2.5%) of positive samples, the molecular method developed in this study proved to be highly sensitive and accurate allowing to conduct an extensive monitoring analysis for these parasites in wildlife.

     

Association of MC3R gene polymorphisms with body weight in the red fox and comparative gene organization in four canids

There are five genes encoding melanocortin receptors. Among canids, the genes have mainly been studied in the dog (MC1R, MC2R and MC4R). The MC4R gene has also been analysed in the red fox. In this report, we present a study of chromosome localization, comparative sequence analysis and polymorphism of the MC3R gene in the dog, red fox, arctic fox and Chinese raccoon dog. The gene was localized by FISH to the following chromosome: 24q24‐25 in the dog, 14p16 in the red fox, 18q13 in the arctic fox and NPP4p15 in the Chinese raccoon dog. A high identity level of the MC3R gene sequences was observed among the species, ranging from 96.0% (red fox – Chinese raccoon dog) to 99.5% (red fox – arctic fox). Altogether, eight polymorphic sites were found in the red fox, six in the Chinese raccoon dog and two in the dog, while the arctic fox appeared to be monomorphic. In addition, association of several polymorphisms with body weight was analysed in red foxes (the number of genotyped animals ranged from 319 to 379). Two polymorphisms in the red fox, i.e. a silent substitution c.957A>C and c.*185C>T in the 3′‐flanking sequence, showed a significant association (P < 0.01) with body weight.