Inappropriate feeding practice favors the transmission of Trichinella papuae from wild pigs to saltwater crocodiles in Papua New Guinea

The recent discovery of Trichinella zimbabwensis in farmed crocodiles (Crocodilus niloticus) of Zimbabwe and its ability to infect mammals, and the development of both T. zimbabwensis and Trichinella papuae in experimentally infected reptiles led to an investigation of Trichinella infection in saltwater crocodiles (Crocodylus porosus) and in wild pigs (Sus scrofa) of Papua New Guinea, to see if T. papuae also, is present in both cold- and warm-blooded animals. Of 222 crocodiles examined, 47 animals (21.2%), all from Kikori, Gulf Province, were positive for non-encapsulated larvae in the muscles. The greatest number of larvae was found usually in the biceps, with an average of 7 larvae/g. One isolate from a crocodile infected successfully both laboratory rats and mice. Of 81 wild pigs examined, 9 from Bensbach river area (Western Province) and 1 from Kikori area (Gulf Province) were positive for non-encapsulated larvae in the muscles. Trichinella larvae from both saltwater crocodiles and wild pigs have been identified by multiplex-PCR analysis as T. papuae. The sequence analysis of the region within the large subunit ribosomal DNA, known as the expansion segment V, has shown the presence of a molecular marker distinguishing T. papuae isolates of Bensbach river area from those of Kikori area. This marker could be useful to trace back the geographical origin of the infected animal. The epidemiological investigation carried out in the Kikori area has shown that local people catch young crocodiles in the wild and keep them in holding pens for several months, before sending them to the crocodile farm in Lae (Morobe Province). They feed the crocodiles primarily with wild pig meat bought at the local market and also with fish. These results stress the importance of using artificial digestion for routinely screening of swine and crocodiles, and of adopting measures for preventing the spread of infection, such as the proper disposal of carcasses and the adequate freezing of meat.     

First record of Trichinella pseudospiralis in the Slovak Republic found in domestic focus

Infection of Trichinella spp. is widespread among wildlife in Slovakia and the red fox (Vulpes vulpes) is the main reservoir of Trichinella britovi. Trichinella spiralis has been rarely documented in sylvatic and domestic animals of this country. During routine examination of domestic pigs at the slaughter, Trichinella larvae were detected by artificial digestion in a domestic pig of a large-scale breeding farm in Eastern Slovakia. The parasite has been identified by molecular (PCR) and biochemical (allozymes) analyses and by the morphology of the nurse cell as the non-encapsulated species Trichinella pseudospiralis infecting both mammals and birds. The epidemiological investigation carried out at the farm level revealed the presence of the same parasite species in other three pigs of 192 examined (2.1%), in 3 of 14 (21.4%) examined synanthropic rats (Rattus norvegicus) and in a domestic cat. The farm was characterized by inadequate sanitary conditions, insufficient nutrition, cannibalism and the presence of rat population. A different profile has been observed at the phosphoglucomutase locus in T. pseudospiralis isolates from Slovakia in comparison with the T. pseudospiralis reference isolate from the Palearctic region. This is the first documented focus of T. pseudospiralis from Central Europe. The detection in domestic pigs of a non-encapsulated parasite infecting both mammals and birds stresses the need to avoid the use of trichinelloscopy to detect this infection at the slaughterhouse.     

Characterisation of Trichinella isolates from Bulgaria by molecular typing and cross-breeding

Trichinella spp. larvae were collected from domestic and wild-life animals in association with 15 human trichinellosis outbreaks registered between 1999-2002 in Bulgaria. Furthermore, Trichinella spp. isolates were obtained from 62 naturally infected wild animals and of a rat. All isolates were subjected to speciation by both multiplex PCR and cross-breeding experiments. Epidemiological and clinical data were collected and analysed using standard protocols for epidemiological surveillance and control of outbreaks. Only two species were identified-Trichinella britovi and Trichinella spiralis. Results obtained by molecular typing fully matched those of cross-breeding. More specifically, parasite isolates obtained upon 15 epidemic outbreaks revealed the predominance of T. britovi (n = 10) when compared to T. spiralis (n = 5). With regard to host origin, the predominant species detected among wild boar was T. britovi (n = 4), and T. spiralis was identified in one wild boar sample only. Among the isolates obtained from domestic pig products, T. britovi was found in five cases and T. spiralis in four cases, respectively. In the naturally infected wild animals not related to epidemics, only T. britovi was demonstrated. The present results provide a strong indication that both T. britovi and T. spiralis operate within domestic and sylvatic cycles in Bulgaria. Geographically, the distribution of T. britovi appears to include Central, Southern, Eastern and Western parts of the country, and wildlife animals from the Mid Balkan Mountains and Mid Sredna Gora Mountains, T. spiralis was found in Western and Southwestern Bulgaria, only.     

Simulating the spread of classical swine fever virus between a hypothetical wild-boar population and domestic pig herds in Denmark

Denmark has no free-range wild-boar population. However, Danish wildlife organizations have suggested that wild boar should be reintroduced into the wild to broaden national biodiversity. Danish pig farmers fear that this would lead to a higher risk of introduction of classical swine fever virus (CSFV), which could have enormous consequences in terms of loss of pork exports. We conducted a risk assessment to address the additional risk of introducing and spreading CSFV due to the reintroduction of wild boar. In this paper, we present the part of the risk assessment that deals with the spread of CSFV between the hypothetical wild-boar population and the domestic population. Furthermore, the economic impact is assessed taking the perspective of the Danish national budget and the Danish pig industry. We used InterSpreadPlus to model the differential classical swine fever (CSF) risk due to wild boar. Nine scenarios were run to elucidate the effect of: (a) presence of wild boar (yes/no), (b) locations for the index case (domestic pig herd/wild-boar group), (c) type of control strategy for wild boar (hunting/vaccination) and (d) presence of free-range domestic pigs. The presence of free-range wild boar was simulated in two large forests using data from wildlife studies and Danish habitat data. For each scenario, we estimated (1) the control costs borne by the veterinary authorities, (2) the control-related costs to farmers and (3) the loss of exports associated with an epidemic. Our simulations predict that CSFV will be transmitted from the domestic pig population to wild boar if the infected domestic pig herd is located close to an area with wild boar (<5 km). If an outbreak begins in the wild-boar population, the epidemic will last longer and will occasionally lead to several epidemics because of periodic transfer of virus from groups of infected wild boar to domestic pig herds. The size and duration of the epidemic will be reduced if there are no free-range domestic pig herds in the area with CSF-infected wild boar. The economic calculations showed that the total national costs for Denmark (i.e. the direct costs to the national budget and the costs to the pig industry) related to an outbreak of CSF in Denmark will be highly driven by the reactions of the export markets and in particular of the non-EU markets. Unfortunately, there is a substantial amount of uncertainty surrounding this issue. If hunting is used as a control measure, the average expenses related to a CSF outbreak will be 40% higher if wild boar are present compared with not present. However, a vaccination strategy for wild boar will double the total costs compared with a hunting strategy.     

Effects of Oesophagostomum dentatum and dietary carbohydrates on morphology of the large intestine of pigs

The effects of Oesophagostomum dentatum infection and dietary carbohydrates on the morphology and epithelial cell proliferation in the gastrointestinal tract of pigs were investigated experimentally. Thirty-two worm-free pigs (n=32) from a specific pathogen-free farm were randomly divided into four groups (A-D), of eight animals each. Pigs in groups A (control) and B (infected) were fed Diet 1, and pigs in groups C (control) and D (infected) were fed Diet 2. The two diets were formulated: Diet 1 (%) contained barley flour, oat husk meal plus soya bean meal (55:21:24) and Diet 2 (%) contained barley flour, inulin and sugar beet fibre (SBF) (80.1:7:12.9) plus soya bean meal (3:1) to contain carbohydrates from inulin and sugar beet fibre (SBF) that were readily fermentable in the large intestine. The two infected pig groups (16 pigs total) were inoculated with 6000 infective larvae of O. dentatum and all pigs, including the controls, were slaughtered 12 weeks p.i. The combination of O. dentatum infection and highly fermentable dietary carbohydrates affected the mucosal architecture, the epithelial cell proliferation and mucin secretion of the large intestine. Infection had a significant influence on the crypt volume, height and density, and on muscularis externa at the proximal and middle colon. The changes in the affected gut sections were proportional to the number of worms present. However, these parameters appeared unaffected by those diets alone. In pigs without infection non-digestible dietary carbohydrates significantly influenced the tissue weight of colon.     

Trichinella pseudospiralis in pig from Croatia

Trichinella spiralis and Trichinella britovi are species that are frequently found in domestic pigs and various sylvatic animals in Croatia. During routine trichinoscopy, non-encapsulated larvae were detected in the muscle tissue of a domestic pig. Artificial digestion revealed a larvae burden of 602 muscle larvae per gram of tissue. Tissue section analysis confirmed the presence of non-encapsulated larvae. Multiplex PCR identified the larvae as T. pseudospiralis. This observation is consistent with the reports of a local veterinary inspector who described the presence of non-encapsulated Trichinella in four individual cases over the last 2 years. This is the first report of T. pseudospiralis in Croatia and one of very few cases of T. pseudospiralis infection described in domestic pigs. The detection of non-encapsulated larvae stresses the need for implementation of artificial digestion instead of trichinoscopy for the detection and identification of Trichinella infections.     

Spatial distribution of Trichinella britovi, T. spiralis and T. pseudospiralis of domestic pigs and wild boars (Sus scrofa) in Hungary

Trichinellosis is a foodborne disease caused by the consumption of raw meat and raw meat-derived products from swine, horse and some game animals infected with nematode worms of the genus Trichinella. Between June 2006 and February 2011, 16 million domestic pigs and 0.22 million wild boars (Sus scrofa) were tested for Trichinella sp. in Hungary. Trichinella infection was not found in any pigs slaughtered for public consumption. Nevertheless, Trichinella spiralis was detected in four backyard pigs when trace back was done following a family outbreak. Trichinella infection was demonstrated in 17 wild boars (0.0077%). Larvae from wild boars were identified as Trichinella britovi (64.7%), T. spiralis (29.4%) and Trichinella pseudospiralis (5.9%). Although the prevalence of Trichinella sp. infection in wild boars and domestic pigs is very low, the spatial analysis reveals that the level of risk differs by region in Hungary. Most of the T. britovi infected wild boars (63.6%) were shot in the north-eastern mountain area of Hungary; whereas domestic pigs and wild boars infected with T. spiralis were detected only in the southern counties bordering Croatia and Romania. In the north-western and central counties, the prevalence of Trichinella infection seems to be negligible.     

IgG response in guinea pigs to Trichinella spiralis infection

An enzyme-linked immunosorbent assay (ELISA) using excretory-secretory antigens was developed to study the dynamics of the IgG antibody response to varying levels of Trichinella spiralis infection in the guinea pig. Four groups of four Hartley guinea pigs each were infected with 1250, 250, 50 or 10 T. spiralis infective muscle larvae. They were bled every 15 days for 6 months and the IgG antibody response determined by ELISA. The time of seroconversion was dose dependent as the larger the dose, the earlier the response occurred. Significant differences in antibody response between the dose groups were evident at 30 days post-infection (P less than 0.05). Beyond 60 days post-infection, the response was similar in the four groups. The antibody response in the groups infected with 250 and 50 infective larvae was similar, but was significantly different from that of the high (1250) and low (10) dose groups from 30 days post-infection (P less than 0.01). Once seroconversion occurred, the antibody titer rose to the same level, irrespective of the initial dose. To compare the antibody response according to muscle larvae recovered, the guinea pigs were grouped into four categories: less than 10 larvae; 10-25 larvae, 50-80 larvae, greater than 100 larvae. A significant positive correlation (P less than 0.05) was observed at 60 days post-infection when these groups were compared.     

Welfare assessment in porcine biomedical research - suggestion for an operational tool

In recent years, increasing interest in using the pig (Sus scrofa) for biomedical research has become evident. Today, the pig is considered an advantageous alternative animal model for various human diseases and conditions. However, even though a considerable amount of biomedical research has been done on pigs, hardly any studies include systematic welfare assessment. Still, it is essential to assess welfare of laboratory pigs, both domestic pig breeds and smaller purpose-bred breeds, as (1) scientific obligations entail responsibility to ensure and document a fair welfare standard for animals used for experimental purposes; and (2) the scientific outcome can be dependent upon the welfare state of the animals. In order to be able to quantify and control laboratory pig welfare, a practical tool is needed. The purpose of the present paper is to provide an overview of the current status of the extent of welfare assessment in pigs used in biomedical research and to suggest a welfare assessment standard for research facilities based on an exposition of ethological considerations relevant for the welfare of pigs in biomedical research. The tools for porcine welfare assessment presented suggest a method for monitoring the welfare status of individual laboratory pigs, intended to relieve the practical scoring of the welfare of individual pigs as well as the interpretation of the findings.     

Toxocara cati larva migrans in domestic pigs - detected at slaughterhouse control in Norway

Routine Trichinella meat inspection at the slaughterhouse detected one larva in a pooled batch of 100 pig samples. The larva was sent to the Norwegian Veterinary Institute (NVI) for species identification.Morphological examination revealed that the larva was not Trichinella spp. Molecular analysis was performed. PCR and sequencing of 5S/ITS identified the larva as Toxocara cati. A second round of digests was carried out at the meat inspection laboratory, in smaller batches to try to identify the infected animal. No further larvae were detected and it was not possible to identify which of the 100 animals the larva had come from. This is the first time that Toxocara cati has been reported in slaughterhouse pigs in Norway.Although the infected individual could not be identified, the meat originated from one of six potential farms. A small survey regarding rodent control and cats was sent to each of these farms. Cats had restricted access to food storage areas (two farms reported that cats had access) whilst none of the farms allowed cats into the production housing. Cats were, however, present on all the farms (mostly stray cats of unknown health status). Half of the farms also reported seeing rodents in the pig housing during the previous six months and half reported finding rodents in the feed and straw storage areas. We were unable to narrow down the source of infection – however contamination of food or bedding material, with cat faeces or infected rodents, in addition to the presence of infected rodents in pig housing remain potential routes of infection.     

银染核仁组成区（Ag—NOR）与家猪品种的起源进化

本文发现Ag-NOR是研究家畜起源进化的一个良好的细胞遗传学指标。它表型稳定,灵敏性高,能够察觉品种间的微小变异。用生物数学方法进行数据处理,x~2独立性检验和遗传距离分析,揭示了中国家猪、欧洲家猪、美洲家猪在起源进化中的相互关系和特点,同时还可对近缘品种加以区分,深化对品种形成过程的认识。     

Trichinosis in a herd of swine: cannibalism as a major mode of transmission

In a herd of approximately 1,000 hogs, evaluation of muscle specimens collected at various intervals during a 12-year period (1973 to 1985) indicated continuous transmission of Trichinella spiralis. The farm's rat population and the incidence of trichinosis in the rats was high during 1974, but diminished markedly by 1978. In January 1984, a longitudinal investigation, using tracer pigs, was performed to determine whether rodents and/or other wild animals were involved in transmission of T spiralis on this farm. Tracer pigs exposed to rodents and wild animals did not become infected with T spiralis. The rodent population on the farm was small and none of the rodents trapped and examined were found to be infected. Hog cannibalism also was evaluated as a mode of T spiralis transmission. Results of the investigation indicated that hog cannibalism was the mode of transmission for trichinosis in the herd.     

Real-time PCR as a surveillance tool for the detection of Trichinella infection in muscle samples from wildlife

Trichinella nematodes are the causative agent of trichinellosis, a meat-borne zoonosis acquired by consuming undercooked, infected meat. Although most human infections are sourced from the domestic environment, the majority of Trichinella parasites circulate in the natural environment in carnivorous and scavenging wildlife. Surveillance using reliable and accurate diagnostic tools to detect Trichinella parasites in wildlife hosts is necessary to evaluate the prevalence and risk of transmission from wildlife to humans. Real-time PCR assays have previously been developed for the detection of European Trichinella species in commercial pork and wild fox muscle samples. We have expanded on the use of real-time PCR in Trichinella detection by developing an improved extraction method and SYBR green assay that detects all known Trichinella species in muscle samples from a greater variety of wildlife. We simulated low-level Trichinella infections in wild pig, fox, saltwater crocodile, wild cat and a native Australian marsupial using Trichinella pseudospiralis or Trichinella papuae ethanol-fixed larvae. Trichinella-specific primers targeted a conserved region of the small subunit of the ribosomal RNA and were tested for specificity against host and other parasite genomic DNAs. The analytical sensitivity of the assay was at least 100 fg using pure genomic T. pseudospiralis DNA serially diluted in water. The diagnostic sensitivity of the assay was evaluated by spiking 10 g of each host muscle with T. pseudospiralis or T. papuae larvae at representative infections of 1.0, 0.5 and 0.1 larvae per gram, and shown to detect larvae at the lowest infection rate. A field sample evaluation on naturally infected muscle samples of wild pigs and Tasmanian devils showed complete agreement with the EU reference artificial digestion method (k-value=1.00). Positive amplification of mouse tissue experimentally infected with T. spiralis indicated the assay could also be used on encapsulated species in situ. This real-time PCR assay offers an alternative highly specific and sensitive diagnostic method for use in Trichinella wildlife surveillance and could be adapted to wildlife hosts of any region.     

Seroprevalence of Toxoplasma gondii in wild pigs (Sus scrofa) from Spain

Sera collected from 507 hunter-killed wild pigs (Sus scrofa) between 1993 and 2004 from five geographic regions in northern Spain and seven regions in southern Spain were assayed for antibodies to Toxoplasma gondii by the modified agglutination test (MAT). Antibodies to T. gondii were detected in 185 (38.4%) of 507 pigs with titers of 1:25 in 71, 1:50 in 111 and > or =1:500 in 3; seroprevalence was significantly higher (P<0.05) in pigs from southern regions. Seroprevalence was density dependent; it was higher in pigs from high stocking per hectare and availability of forage. Statistically significant differences were not observed between T. gondii seroprevalence and hunting estates (open versus fenced), sex or age. Serological results indicate a widespread exposure to T. gondii among Spanish wild boars, suggesting that this population could represent a public health risk for persons that handle or consume raw or undercooked infected wild pig meat.     

A Slaughter Slab Survey for Extra-Intestinal Porcine Helminth Infections in Northern Tanzania

A survey on extra-intestinal porcine helminths was conducted at three slaughter slabs that receive pigs from Mbulu, a district endemic for porcine cysticercosis in northern Tanzania. Seventy carcases of pigs between 1 and 2 years old were examined between December 1997 and March 1998. The examination involved ante-mortem lingual examination for Taenia solium cysticercosis followed by post-mortem inspection. In addition, a laboratory procedure was performed to determine whether any of these domestic pigs were infected with Trichinella species. Parasites detected were Ascaris suum (44.3%), Echinococcus granulosus (4.3%) and Taenia hydatigena (1.4%). The lack of cases of porcine cysticercosis in this study compared to previous studies suggests that pig traders are conducting their own ante-mortem lingual examinations before purchasing pigs in the rural communities where the parasite is still highly prevalent. It is concluded that improved meat inspection could prove useful in reducing the local population's risk of infection with these parasites. The results of this study have revealed the parasites of agricultral and public health importance in the targeted communities. Further epidemiological investigations are required to better determine parasite prevalence and impact in order to formulate appropriate and cost-effective strategies for control.     

Molecular epidemiology of classical swine fever in Italy

To gain an insight into the molecular epidemiology of classical swine fever (CSF) in Italy, virus isolates originating from outbreaks that occurred between 1985 and 2000 in wild boar or in domestic pigs in mainland Italy and in Sardinia were analysed by genetic typing. For this, a fragment (190 nucleotides) of the E2 glycoprotein gene was sequenced and phylogenetic analyses were performed, including older Italian isolates and isolates from recent outbreaks in Europe for comparison. The results show that in mainland Italy, several independent epidemiological events occurred in the last decade. In the north of the country, viruses of genotype 2.2 have persisted in wild boar, causing sporadic outbreaks in domestic pigs. In contrast, viruses of subgroups 2.1 and 2.3 appeared only intermittently in different regions of the mainland. In 1997, classical swine fever virus (CSFV) isolates belonging to the subgroup 2.1 and genetically and epidemiologically related to the Dutch isolate in Venhorst, affected domestic pigs exclusively. The isolates of subgroup 2.3, derived from wild boar as well as from domestic pigs were closely related to isolates from Germany and Poland. In Sardinia, CSF is an endemic in wild boar and affects domestic pigs also. Genetic typing showed that viruses of subgroups 1.1 and 2.3 have been present, the last ones being unrelated to the mainland viruses of the same subgroup. Due to the large quantities of pig and wild boar meat imported in some parts of Italy, it cannot be established if these viruses were always present in either the mainland or Sardinia, or if they represent recent introductions.     

Comparative assessment of a double antibody enzyme immunoassay test kit and a triple antibody enzyme immunoassay for the diagnosis of Trichinella spiralis spiralis and Trichinella spiralis nativa infections in swine.

Enzyme immunoassays using the triple antibody enzyme linked immunosorbent assay (ELISA) with both Trichinella spiralis spiralis and T. spiralis nativa excretory-secretory (ES) antigens and a commercial Trichinella spiralis enzyme immunoassay test kit were carried out on sera from pigs that were infected with light, moderate and high doses of infective T. spiralis spiralis and T. spiralis nativa respectively. Seroconversion occurred in all pigs given infective Trichinella larvae although no trichinae were recovered from pigs given T. spiralis nativa larvae and examined between days 92 and 99 postinfection by pepsin digestion. Anti-Trichinella antibodies were detected in pigs infected with T. spiralis spiralis and T. spiralis nativa by ELISA using either the homologous or heterologous ES antigen. The commercial Trichinella spiralis enzyme immunoassay test kit also detected anti-Trichinella antibodies in both the T. spiralis spiralis and T. spiralis nativa infected pigs. The commercial test kit did not appear to be as sensitive as the triple antibody ELISA since it usually took two to three days longer for seroconversion to be detected by the former procedure. Finally seroconversion occurred more rapidly in swine infected with T. spiralis spiralis than with pigs receiving comparable doses of T. spiralis nativa.     

Detection of Trichinella spiralis nativa antibodies in porcine sera by ELISA using T. spiralis spiralis excretory-secretory antigen

Enzyme-linked immunosorbent assay examination of sera from pigs vaccinated with T. spiralis nativa infective larvae and/or challenged with T. spiralis spiralis larvae using a T. spiralis spiralis excretory-secretory antigen showed a significant cross-reaction between the two species of Trichinella. Eight of 12 pigs vaccinated with a high dose of T. spiralis nativa reacted positively 28 days postvaccination while the remaining four pigs had high but negative ELISA optical density readings. Five of six pigs challenged with the homologous species reacted positively 28 days postchallenge but the sixth pig remained negative despite having a muscle infection of 5.6 larvae/g of musculature.     

Methods for investigating Trichinella infections in domestic and wild animals

Trichinellosis is an important parasitic zoonosis that is caused by the intracellular nematode Trichinella spp.. Infection of humans occurs through consumption of raw (or undercooked) meat containing infectious larvae. In Europe, meat from pork, horse, and wild boar have been identified as most important sources of Trichinella infections in humans. In Switzerland, both the domestic pig and wild boar population are considered free of Trichinella. Conversely, Swiss foxes, lynxs and recently a wolf were found to be infected, the species identified in these animals was always referred to as Trichinella britovi. Although this species rarely infects pork and, compared to Trichinella spiralis, only causes reduced pathogenic effects in humans, the basic presence of Trichinella in Switzerland cannot be neglegted. This fact has gained increasing importance since the responsible authorities in the European Union (EU) are preparing regulations for the official Trichinella-control in meat in order to improve food safety for consumers. These regulations will be implemented as a consequence of the recent association of east European countries with the EU. This new legislation particularly takes into account, that in the past by far most cases of human trichinellosis in the EU were due to consumption of imported east European meat.Within the framework of the bilateral agreements of Switzerland with the EU, the Swiss veterinary public health authorities will have to comply with the foreseen EU regulations. Although diagnostic methods for the direct demonstation of Trichinella in pork meat are already routine practice in several Swiss abattoirs, the implementation of a meat control program for Trichinella for the entire slaughter pig population of the country would lead to an enormous increase in costs for the administration and will require an increased infrastructure in veterinary services. In order to find a reduced testing format for monitoring Trichinella infections in Swiss pork, an infection risk-oriented survey strategy is currently evaluated. In the present article, this minimized survey strategy is discussed regarding its compatibility with the EU regulations laying down rules for the official control of meat for Trichinella.     

二花脸猪的高分辨显带核型研究

采用MTX或TdR细胞同步化技术制备了早中期染色体,并应用G-,R-,C-,DA/DAPI显带和银染技术,研究了二花脸猪的高分辨显带核型,其主要结果如下:1.G-带和R-带带型与其它家猪的带型相似,但C-带和Ag-NORs存在多态性;2.Ag-NOR定位于No.7和No.10染色体上;3,DA/DAPI带与端着丝粒染色体C-带相对应。此外,绘制了G带核型模式图,并讨论了染色体的分组及排列、G带带型的一致性、着丝粒异染色质的异质性和Ag-NOR频率与品种的关系。