I. Antiserum Production

These studies report on the production of African swine fever antiserum for use in serological tests. The first attempt to obtain antiserum was made by inoculating ASF virus - infected pig blood into the lactiferous sinus of lactating bovines. This failed to result in the development of detectable antibody, but resulted in propagation of the virus over a 14 to 21 day period.

In the second attempt use was made of a tissue culture - attenuated virus to produce resistance in normal pigs. Clinical response to inoculation with the attenuated virus was limited to a one day increase of temperature. These pigs were subsequently orally exposed to virulent ASF virus and later challenged by intramuscular injection. The sera were subjected to testing by the modified direct complement-fixation test and the agar gel double-diffusion technique in order to follow the development of antibodies. Some sera were also conjugated with fluorescein isothiocyanate and used for the detection of viral antigen by the fluorescent antibody technique.

It was found that inoculation with the attenuated virus brought about the development of low antibody levels in the pigs. This antibody level did not increase following oral exposure. One pig following intramuscular challenge underwent a series of ascending temperature peaks, coinciding with increased complement-fixing titres.

     

Concentrations of antisecretory factor in eggs and in chicken blood plasma

1. In the present study, the concentration of antisecretory factor (ASF), a lectin with hormone‐like properties, and with the capacity to regulate water and electrolyte transport in the small intestine, was higher in hen's egg yolk than in egg white (1·20 v. 0·46 units/ml).

2. The blood plasma concentrations of ASF activity were higher in d‐old chicks (1·03 units/ml) than in 7‐d‐oId (0·42 units/ml) or 21‐d‐old birds (0·18 units/ml); the contents were found to be high again in 35‐d‐old chickens, but to have decreased by 60 to 90% after transport to slaughter 1 day later.

3. The amount of ASF activity was lower in two groups of chickens manifesting patent loose droppings at slaughter (0·15 units/ml in one group, and 0·08 units/ml in the other), than in two groups with normal faecal consistency (0·65 units/ml in one group, and 0·72 units/ml in the other).

4. The results demonstrate the presence of ASF in eggs, and a variation of ASF activity in chickens blood plasma in relation to age, stress and faecal consistency. The interpretation of these data suggests a regulatory influence of ASF on chicken intestinal transport of water.     

Sanitary measures in piggeries,awareness, and risk factors of African swine fever in Benue State,Nigeria

The present study describes assessment of sanitary measures in piggeries of Benue State, Nigeria, to identify the risk factors of African swine fever. Questionnaires were distributed to 74 respondents consisting of piggery owners and attendants in different piggeries across 12 local government areas (LGAs) to collect data for this study. Sanitary measures in piggeries were observed to be generally very poor, though respondents admitted being aware of ASF. Piggeries located within 1-km radius of a slaughter slab (OR?=?9.2, 95% CI 3.0–28.8; p?<?0.0001) and piggeries near refuse dump sites (OR?=?3.0, 95% CI 1.0–9.5; p?<?0.05) showed higher chances of African swine fever virus (ASFV) infection, while piggeries where farm workers wear their work clothes outside of the piggery premises (OR?=?0.2, 95% CI 0.1–0.7; p?<?0.01) indicate less chances of infection but had a significantly associated p value thus were identified as potential risk factors. The study concluded that pigs in Benue State are still at risk of an ASF outbreak. Proper sanitary and hygienic practices are advocated and emphasized in piggeries, while routine surveillance for African swine fever virus antibodies in pigs in Benue State is strongly recommended to provide a reliable reference database to plan for the prevention of any devastating ASF outbreak.

     

Pathogenesis of Rabies in the Rat

Three strains of rabies virus, one fixed and two street, were compared for invasiveness in neurectomized rats following foot pad inoculation. Approximately one-fifth of test rats studied revealed that the virus bypassed the severed sciatic and saphenous nerves and reached the brain.

All rats dying within 20 days were positive for rabies upon fluorescent antibody and mouse mortality tests.

Viremia was demonstrated in 20 per cent of both neurectomized or non-neurectomized rats one hour after inoculation of rabies virus, both fixed and street strains, into the foot pad.

     

II. Detection of the Virus in Swine Tissues by Means of the Modified Direct Complement-Fixation Test

The modified direct complement-fixation test, supplemented with unheated normal calf serum, was used to demonstrate antibodies in sera of swine immunized to African swine fever virus. These antibodies did not react in the ordinary direct non-supplemented complement-fixation test.

African swine fever complement-fixing antigen in infected swine tissue is not denatured by extraction with fat solvents. Consequently, good antigens devoid of non-specific reactivity were obtained by extraction with a mixture of acetone and ether.

The virus was detected in infected swine tissue harvested one day after beginning of pyrexia. The modified direct complement-fixation test demonstrated cross-reactions between the six strains of virus studied.

     

非洲猪瘟自然弱毒株演变进程

自2018年非洲猪瘟(African swine fever,ASF)传入中国以来疫情得到了有效控制。但2021年以后,我国的ASF流行情况出现了新的变化,临床中出现了“自然变异株”。这些毒株可能导致感染后临床症状不典型,容易与其他疫病混淆等问题。这与国外的流行演变规律一致,即当ASF在一个国家或地区流行较长时间后,其临床表现将由急性发病转变为缓慢发病或出现新的临床表现。经过多年的观察积累,国外已经对该病毒的流行规律特别是自然弱毒株的演变进程有了大量记载,但国内目前尚无这方面的描述。为此,就ASF自然弱毒株的演变进程进行综述,从而提出加强血清学诊断产品研发、加快产业化报批进程、适时开展血清学监测追溯等建议,以期为我国ASF科学防控提供参考。     

Investigation into the pathogenesis of Atrophic Rhinitis in pigs

Summary

In two groups of swine herds, herds with and without clinical AR the presence of Atrophic Rhinitis (AR) correlated with the presence of toxinogenic Pasteurella multocida (PM) and not with the Bordetella bronchiseptica (BB) infection.

Six BB‐ and eighteen PM‐strains have been investigated for AR pathogenicity. Broth cultures were injected intradermally in guinea‐pigs (GPST) orintranasally in 3‐week‐old colostrum deprived specific pathogen free (SPF) piglets.

The average atrophy of the ventral conchae (A VC) correlated with the GPST in 4BB‐ and 7 PM‐strains. One BB‐ and 2 PM‐strains were qualified as doubtful, the others as non‐AR pathogenic. With AR pathogenic BB‐ and PM‐strains clinical AR could be induced in 3‐ and 6‐week‐old piglets. AVC lesions (gradation> 1) could be induced with BB in piglets of 6 and with pathogenic PM in 16‐week‐old piglets. Six of seven AR pathogenic PM‐strains resembled Carter‐type D and one resembled type A. No significance was found between AR pathogenicity and somatic serotypes.

Intranasal instillations of cell‐free broth culture filtrates of AR pathogenic PM‐strains also caused AR in piglets. These filtrates also caused lethality in piglets and in mice lethalitytest (MLT) and induced a positive GPST. After heating the pathogenic effects of the filtrates disappeared. The name AR toxin has been introduced for this thermolabile, haemorrhagic dermonecrotic (HDNT) fraction of the AR inducing filtrates. The severity of the AR lesions depended on the amount of the AR toxin intranasally instilled in pigs.

Cross protecting antibodies obtained in rabbits against the AR toxins of two PM strains could be demonstrated by a toxin neutralisation test in the MLT and the GPST.

Broth cultures were injected intradermally in guinea‐pigs (GPST) or intranasally in 3‐week‐old colostrum deprived specific pathogen free (SPF) piglets.     

The Significance of Proliferation and Enterotoxin Production by Escherichia coli in the Intestine of Gnotobiotic Pigs

The significance of enterotoxin production and proliferative ability of Escherichia coli in the intestinal tract as related to porcine enteric colibacillosis was studied in 68 gnotobiotic pigs.

The animals were monocontaminated at seven to ten days of age with eight selected strains of E. coli. The strains were two naturally occurring porcine enteropathogens — P155 (0149:K91;K88a,c:H10) and P307 (08:K87;K88a,b:H19), two nonenteropathogenic strains — P104 (0139:K82:H1) and F11 (018-ab:K?:H14), and four enterotoxigenic derivatives of the above strains — P104(P155), P104(P307), F11(P155) and F11(P307). The response of the animals was evaluated on the basis of clinical observations and necropsy lesions 22 hours after exposure to the organisms. E.coli counts were determined at seven different levels of the intestinal tract. Cell free extracts of the intestinal contents were examined for enterotoxic activity by the ligated pig intestine loop test.

All of the strains possessing the enterotoxin plasmid produced enterotoxin in the pig's intestine and were capable of causing diarrhea. The nonenteropathogenic E. coli failed to do so. The strains possessing the P155 enterotoxin plasmid were more virulent than the corresponding derivatives with the P307 enterotoxin plasmid. Strains P155, P307 and P104(P155) proliferated in the upper small intestine at a greater rate and were more virulent than the other strains. The numbers attained in the upper small intestine by the other enterotoxigenic derivatives were comparable to those of their nonenteropathogenic parent strains.

It was considered that enterotoxin produced by E. coli was the essential factor for causing a diarrheic response in gnotobiotic pigs. The virulence of each of the tested strains appeared to be governed by the degree of enterotoxicity associated with a particular enterotoxin plasmid, the numbers attained by these organisms in the upper small intestine, (but not in the lower small intestine or in the colon), and by other undetermined factors.

     

African swine fever virus infection of the bushpig (Potamochoerus porcus) and its significance in the epidemiology of the disease

Warthog (Phacochoerus aethiopicus), giant forest hog (Hylochoerus meinertzhageni) and bushpig (Potamochoerus porcus) are known to be susceptible to infection with African swine fever (ASF) virus. Little however, is known about the ecology of the disease in the bushpig. This study has shown that the bushpig remains viraemic for between 35 and 91 days following infection during which time it is able to infect the tick vector O. moubata. These ticks were able to transmit the disease to pigs. The virus persists in the lymphatic tissues for less than 34 weeks. Bushpigs infected with LIL 20/1 virus but not VIC T90/1 virus transmitted infection to in-contact pigs. Infected domestic pigs did not transmit the infection to in-contact bushpigs. ASF virus was able to replicate in in vitro cultures of bushpig leucocytes and endothelial cells. Recovered bushpigs could be re-infected with some strains of virus but not others. While it has been demonstrated that bushpigs remain carriers of ASFV following infection a complete understanding of their significance in the epidemiology of the disease awaits further investigations of their association with O. moubata.     

Bovine Infectious Keratoconjunctivitis: Serological Aspects of Moraxella bovis Infection

A modification of a gel diffusion precipitin test (GDPT) was used to detect antibodies for Moraxella bovis (M. bovis) in the sera of cattle affected with bovine infectious keratoconjunctivitis (BIK). The test was also used for the detection of sequential antibody development in cattle vaccinated with cultures of M. bovis. Also, strains of M. bovis isolated from cattle herds affected with BIK were characterized serologically as a part of an identification scheme using the test.

A comparison of the antigenic properties of various strains of M. bovis and M. bovis-like organisms was conducted using the test. The results indicated that there might be antigenic relationships between M. bovisand M. bovis-like organisms such as Moraxella liquefaciens, Moraxella nonliquefaciens, an unidentified hemolytic diplococcus, Mima polymorpha, Mima polymorpha var. oxidans and Herellea vaginicola

The authors suggest that the GDPT can be used for serological studies of BIK, and the identification and antigenic analysis of M. bovis. They indicate, however, that a more definitive study is needed to evaluate the reliability of the test for quantitative work.

     

非洲猪瘟背景下生猪产业链价格波动及传导研究——以河北省为例

非洲猪瘟疫情的暴发对我国生猪产业,尤其是生猪产业链价格产生巨大影响。为研究非洲猪瘟背景下河北省生猪产业链价格波动及传导机制,文章选取河北省2018年8月至2020年6月周度平均仔猪价格、生猪价格和猪肉价格作为生猪产业链价格代表,通过构建协整和误差修正模型、格兰杰因果检验方法分析非洲猪瘟对生猪产业链内部价格传导过程的影响。研究结果表明：非洲猪瘟背景下河北省生猪产业链价格存在长期均衡关系,但其上下游价格传导受阻,而屠宰场和销售市场的垂直整合度增强;生猪产业链中存在一定的反向误差修正机制;生猪产业链上、中游对下游价格的影响更为明显,并据此提出非洲猪瘟背景下河北省调控生猪产业链价格的政策建议。     

Variability in concentrations of zinc in serum and feed when using zinc oxide as a supplement for the prevention of facial eczema

AIMS: To determine the variability of concentrations of Zn in feed, when used as a supplement to prevent facial eczema, and to determine the variability in concentrations of Zn in serum between cows and herds that are being supplemented with ZnO in feed, using in-shed feeders or on a feed pad.

METHODS: Sixteen commercial dairy farms in the Waikato region of New Zealand were enrolled, that were supplementing cows with ZnO in the feed using either an automatic in-shed feeder (ASF) or a feed pad (FP) using a feed-out or mixer wagon. On each farm 10 cows were selected by the farmer, that were assumed to be representative of the age and liveweight of the herd. Four hours after supplement feeding, each cow was weighed and a blood sample collected for measurement of concentrations of Zn in serum. Three samples of feed were collected from each farm for Zn analysis, from the beginning, middle and end of the feed being distributed. Levene’s test for homoscedasticity was used to analyse whether there were differences in variation of individual concentrations of Zn in serum, and in the feed, between the two feeding systems. Multivariable linear regression was used to examine associations between age, feeding method or liveweight and concentrations of Zn in serum, after accounting for the variability between farms.

RESULTS: Of the 163 cows sampled, concentrations of Zn in serum were between 20–35?µmol/L in 75/163 (46 (95% CI=38–54)%) cows; were <20?µmol/L in 71/163 (44 (95% CI=36–52)%) cows, and >35?µmol/L in 17/163 (10 (95% CI=6–16)%) cows. The variation in concentrations of Zn in serum in individual cows differed between farms (p<0.001), and the variability was greater for cows fed using a FP than ASF (p<0.001). There was no difference in the variation of concentrations of Zn in feed between the two feeding methods (p=0.54), but concentrations of Zn in serum were associated with the amount of Zn offered in feed (p=0.008).

CONCLUSIONS AND CLINICIAL RELEVENCE: There was significant variability between farms in the concentrations of Zn in the serum of cows being supplemented with ZnO in feed. Only 46% of cows sampled had concentrations of Zn between 20–35?µmol/L. Effective management of facial eczema should include monitoring Zn in the feed and in serum to ensure cows are receiving the correct dose they require.     

African Swine Fever: Application of Immunoelectroosmophoresis for the Detection of Antibody

Thirty-three pigs in three groups of nineteen, ten, and four pigs were infected with three different African swine fever (ASF) virus isolates, respectively. All virus isolates were attenuated to varying degrees by passaging in cell cultures, and they retained sufficiently low virulence to produce subacute and chronic infections in pigs. Sera collected at various intervals were tested for antibody activity by the immunoelectroosmophoresis, agar gel diffusion precipitin, and complement-fixation tests using a modified Kolmer technique. Results clearly indicated that the immunoelectroosmophoresis test is a rapid (30 minute) and accurate method with extreme sensitivity and superior to the complement-fixation and agar gel diffusion precipitin tests in detecting antibody against ASF virus. Possible use of this method in detecting ASF virus infection is suggested.     

非洲猪瘟诊断制品注册评审现状分析

2018年8月，非洲猪瘟在我国沈阳首次爆发，随后蔓延至全国，给我国养猪业造成了重大经济损失。利用诊断制品对猪群非洲猪瘟感染情况进行检测与早期诊断，是预防和控制非洲猪瘟的有效手段。2019年以来，非洲猪瘟诊断制品注册申报数量居高不下，但常因申报资料存在各种问题未通过注册评审。鉴于此，对我国非洲猪瘟诊断制品注册申报情况、评审评价政策以及不符合注册要求的原因进行了分析，并提出了相关建议，供非洲猪瘟诊断制品研发、注册人员参考。     

The Effect of Chlortetracycline Feed Additive on Experimental Salmonella Infection of Swine and Antibiotic Resistance Transfer

Four Salmonella strains were used. With two, S. canada and S. enteritidis, establishment of infection in pigs was poor. Although improved establishment was achieved with two strains of S. typhimurium, no infections were extensive. No significant difference was detected between excretion of infecting strains by pigs receiving chlortetracycline at levels of ten gm per ton of feed and that by control counterparts. Comparable establishment of S. typhimurium S192 was achieved in pigs receiving 20 or 40 gm of chlortetracycline per ton.

Resistant variants were isolated from some animals in all experiments using S. typhimurium S192. Almost without exception such variants were isolated only after specimen enrichment. The numbers of specimens from test animals yielding resistant variants were more than twice those of control counterparts. Numbers of such variants in material from the former were much greater.

In vitro transfer of resistance to S. typhimurium S192 was demonstrated both in waters from drinking troughs and during specimen enrichment. The former, indicating the risk of transfer in pigs' environments and packing houses, has disturbing implications for public health. The latter emphasizes the difficulties of determining antibiotic sensitivity of pathogens as they occur in the animal body.

     

Laboratory diagnosis and disease occurrence in the current African swine fever eradication program in Spain, 1989–1991

The purpose of this study was to determine the rate of decline and geographical distribution by municipality of clinical and subclinical African swine fever (ASF) in the affected areas of Spain. A second aim was to evaluate the performance of diagnostic tests in the Spanish ASF eradication program. Clinical outbreaks were confirmed using both the direct and indirect immunofluorescence test (and if both were negative, by the hemabsorption test). The serological status of swine was determined by an enzyme-linked immunosorbent assay (ELISA) and suspect serum samples were confirmed by the immunoblot assay.

The number of clinical outbreaks (herds) of ASF for 1989, 1990 and 1991 was 170, 347 and 207, respectively. The numbers of municipalities within each affected province experiencing acute outbreaks for the same time periods were 49, 69 and 48, respectively. Serologically diagnosed animals positive for ASF were 1.1% of animals tested in 1989, 0.5% in 1990 and 0.8% in 1991. The corresponding positive predictive values of the standard ELISA test used were 99.0, 97.9 and 98.8, respectively. Similarly, the number of municipalities within each affected province experiencing serologically positive subclinically infected animals was 269, 178 and 147 for each of the years 1989, 1990 and 1991, respectively.     

V. Cultivation of the Virus in Primary Pig Kidney Cells

Six strains of African swine fever (ASF) virus were propagated in culture of primary pig kidney (PK) cells. The course of virus growth was followed by means of the fluorescent antibody staining technique. All 6 strains multiplied in the cultures, and 5 of these eventually showed cytopathic effects leading to cell death. Three of the strains were tested for pathogenicity in pigs at various passage levels. Each showed evidence of modification in virulence after a relatively few passages in PK cells. In one case modified virus produced resistance to challenge with homologous virulent virus. All strains rendered the PK cultures capable of hemadsorption of pig erythrocytes.     

Terminal Dissemination of Rabies Virus in Selected Rat Tissues

Fifty rats were divided into 3 groups and challenged via the foot pad route with a fixed and 2 different strains of street rabies virus in order to study the dissemination of the virus and the affinity for certain tissues of the rat.

The incubation period for rats inoculated with fixed rabies is shorter than with street virus, being 5 to 7 days compared with 10 to 12 days.

Rats inoculated with the fixed strain were less aggressive and irritable than rats inoculated with street virus.

The fixed strain demonstrated a greater affinity for the tissues studied as compared to the street strains of virus.

Both the fixed and street strains revealed a low affinity for the parotid gland since no virus could be demonstrated in 14 of 20 in the fixed virus group and 27 of 30 in the street virus group.

     

试论中兽医中药对生猪复养工作的重要作用

祖国的中兽医中药学博大精深,近年来在防治非洲猪瘟中中兽医中药发挥了重要作用。文章就"治未病"、"正气存内,邪不可干"中兽医辨证施治和"君臣佐使"处方原则,配制防治非洲猪瘟的中药处方。对中药防治疫病特点、抗病毒的成分、作用机理、中药与西药不同点及各地运用中药防治非洲猪瘟情况,进行了汇总。介绍了湖南省洞口县生猪复养工作开展情况,筛选了3个防治非洲猪瘟中药处方作为洞口县生猪复养技术支撑。收集了治疗登革热等4例中药抗病毒病例和防治非洲猪瘟中药方制剂(药品)10个。提出整合和筛选防治非洲猪瘟中药处方的地方资源,验证和试验中药防治非洲猪瘟效果,尽快研制出有效地防治非洲猪瘟的中药复方制剂药品。     

非洲猪瘟病毒微芯片荧光PCR快速检测技术的建立及初步应用

非洲猪瘟缺乏安全有效的疫苗,其防控有赖于及时、准确的诊断和消毒灭源.本研究在对非洲猪瘟病毒(Afri-can swine fever virus,ASFV)B646L基因序列进行分析的基础上设计引物、探针,建立了基于微芯片的ASFV免提取荧光PCR检测技术.特异性试验结果显示,微芯片PCR可用于我国不同地区ASFV毒株...