混合感染中鸡毒支原体和副鸡嗜血杆菌的分离鉴定

从冀南地区某养鸡场患呼吸道疾病的病鸡群采集5份样品（气管和眶下窦分泌物）中，同时分离到鸡毒支原体和副鸡嗜血杆菌两种病原体。通过病原的分离培养、细菌L型检验、生化试验、血清学试验等鉴定，证明分离的鸡毒支原体与国际标准株S6的血清型一致，副鸡嗜血杆菌的血清型为A型，动物试验表明，分离的鸡毒支原体和副鸡嗜血杆菌均具有明显的致病性，说明该病鸡群同时混合感染了鸡毒支原体和副鸡嗜血杆菌。     

秋冬交替季节鸡常见病的防治

一鸡传染性鼻炎1病原及流行特点本病是由副鸡嗜血杆菌引起的一种鸡的急性上呼吸道传染病。副鸡嗜血杆菌属于革兰氏阴性菌,有A、B、C三个血清型。前几年,我国流行的血清型主要是A型和零星C型,但近年来主要血清型是A和B型,且A型已经出现变异,发病比例有明显增加的趋势。     

预防B型变异株的一种新型传染性鼻炎四价疫苗的效力

传染性鼻炎是由副鸡嗜血杆菌引起的鸡的一种急性呼吸道疾病，目前在生产上主要使用含有代表A、B、C三种血清型（按Page分类（1962））的两种或多种菌株制成的灭活疫苗。到目前为止，这些疫苗对特定血清型菌株的合并感染提供了令人满意的免疫保护。免疫失败通常与某些商品疫苗中遗漏其中些分离株进行血清分型。结果发现了多     

B型副鸡嗜血杆菌的分离鉴定

从辽宁某公司鸡场疑似鸡传染性鼻炎的病鸡眶下窦分离到一株副鸡嗜血杆菌，用Page程序和Kume程序对其进行血清型鉴定，确认为B型副鸡嗜血杆菌，这是首次在我国分离到B型副鸡嗜血杆菌。     

鸡传染性鼻炎(IC)的诊断与防治

<正> 鸡传染性鼻炎(IC)是由副鸡嗜血杆菌(HP)引起鸡的一种急性呼吸道疾病。该菌为革兰氏阴性的多型性小杆菌。上世纪30年代研究认为,IC 病原体的生长需要 X因子(血红素晶)和 V 因子(烟酰胺腺嘌啉二核苷酸),所以被划为嗜血杆菌,直到1962年研究发现所有 IC 病原体的生长都只需要 V 因子,并命名为副鸡嗜血杆菌。众所周知,IC 病原体含有 A、B、C 三个血清型,各型之间无交叉反应。近年来在北美、南美和非洲相继发现副鸡嗜血杆菌的 B 型变异株,此变异株与传统的血清型之间少有交叉反应。鸡传染性鼻炎(IC)可感染各种年龄的鸡,自然条件下多见于育成鸡和成年鸡,尤以产蛋鸡居多。本病一年四季均可发生,但以秋冬及早春季节多发。传播方式以呼吸     

副鸡嗜血杆菌人工感染鸡抗体动态的研究

本研究采用阻断ELISA（B－ELISA）和血凝抑制试验（HI）方法,检测了鸡体分别人工感染副鸡嗜血杆菌Hp8株（A型）和H668株（C型）后0－9周的血清抗体消长情况,并绘制了示意抗体曲线。结果表明,Hp8人工感染后,从第2周到第9周,B－ELISA阳性检出率一直保持100％,其中抗体效价在感染后第4周达到效价高峰,平均约为15．2。而H688人工感染鸡则在感染后第7周检出率最高,此后急剧下降。AI方法对A型抗体的阳性检出率亦在在第4－5周达到高峰,而对C型抗体的检出率一直较低。结果进一步显示了B－ELISA良好的特异性和敏感性,为本方法及其试剂盒产品的进一步应用提供了参考。     

鸡传染性鼻炎的诊断

鸡传染性鼻炎是由细菌引起的一种以面部肿胀、流鼻涕为特征的急性上呼吸道传染病,本病病原副鸡嗜血杆菌通常分为A、B、C 3个血清型[1],且3型均可致病.迄今为止,在我国鸡群中尚未发现有B型副鸡嗜血杆菌感染的证据,多年来,A型副鸡嗜血杆菌一直是我国鸡传染性鼻炎的主要致病菌型,而由C型菌引起的鸡传染性鼻炎少见报道.1998年冬,山东某鸡场发生了一起以肿头为特征的疾病,经作者诊断,认为是鸡传染性鼻炎,而且可能由C型副鸡嗜血杆菌引起.有关情况报告如下.     

鸡传染性鼻炎平板染色诊断抗原的制备及应用

应用鸡副嗜血杆菌（HPG）A型（HPG221）和C型（HPG668）鸡传染性鼻炎（IC）的致病性血清型国际标准株,制备了IC平板凝集试验（SPA）染色诊断抗原。该抗原特异性强：不但与鸡新凤（ND）、鸡白痢、鸡支原体等9种鸡血清无交叉反应,而且可以进行鸡传染性鼻（IC）血清型的鉴定;敏感性高;SPA较琼脂免疫扩散试验（AGID）高2个滴度;稳定性好：4℃保存2年,检测效果不变;应用此抗原进行平板凝集反应检测IC抗体,操作简便,检测快速,适于普及。     

蛋鸡产蛋期传染性鼻炎的诊治

鸡传染性鼻炎是由副鸡嗜血杆菌引起的鸡的一种急性呼吸道疾病。它属于革兰氏阴性菌,该病主要有A、B和C三个血清型;B型为A型的一个变种,即无致病力;而中     

蛋鸡产蛋期传染性鼻炎的诊治

鸡传染性鼻炎是由副鸡嗜血杆菌引起的鸡的一种急性呼吸道疾病。它属于革兰氏阴性菌,该病主要有A、B和C三个血清型;B型为A型的一个变种,即无致病力;而中     

Characterization and use of monoclonal antibodies to identify Haemophilus paragallinarum serovars

Two serovar-specific monoclonal antibodies (MAbs) to Haemophilus paragallinarum serovars A/1 and C/2 strains, respectively, were developed and characterized by hemagglutination-inhibition (HI) and dot-blotting tests using representative H. paragallinarum serovars A/1, B, and C/2 strains. In both the HI and dot-blotting tests, one MAb (E5C12D10), raised against strain 221, serovar A/1, reacted only with serovar A/1 strains, while the other MAb (F2E6), raised against strain S1 of serovar C/2, reacted with only serovar C/2 strains examined. In both tests, the two MAbs did not react with two serovar B strains. These results indicated that the two MAbs recognize serovar-specific hemagglutinating (HA) antigens of H. paragallinarum serovars A/1 and C/2 strains, respectively, and that a dot-blotting test using these MAbs is a practical alternative to the HI test for serotyping H. paragallinarum. Strains 0222 and Spross of serovar B, which did not react with these two MAbs, were found to possess serovar-specific HA antigen in cross-HI tests.     

Characterization of two new monoclonal antibodies against Haemophilus paragallinarum serovar C hemagglutinating antigen

Two new monoclonal antibodies (MAbs), D6D8D5 and B3E6F9, both directed against Haemophilus paragallinarum serovar C hemagglutinating (HA) antigen, were produced, and characteristics of the MAbs were compared with those of the previously described MAb F2E6 in dot-blot and hemagglutination-inhibition (HI) tests using two representative H. paragallinarum strains each of serovars A, B, and C strains and 55 Japanese serovar C field isolates. MAb D6D8D5 and MAb F2E6 reacted with all serovar C strains and field isolates in the dot-blot test. However, MAb D6D8D5 showed various degrees of inhibition of the HA activity of field isolates. In the enzyme-linked immunosorbent assay-competition test, MAb D6D8D5 did not compete with MAb F2E6. MAb B3E6F9 reacted with strain S1, serovar C but not with strain Modesto, serovar C in both dot-blot and HI tests. Three out of 55 field isolates did not react with MAb B3E6F9. Neither MAb reacted with the serovar A and B strains.     

Evaluation of a panel of monoclonal antibodies in the subtyping of Haemophilus paragallinarum.

A panel of four monoclonal antibodies (MAbs) was evaluated, using a hemagglutination-inhibition test, for its ability to subtype 76 isolates of Haemophilus paragallinarum. The results of the MAb reactions were compared with the results of both the Page and Kume serotyping schemes (the serovars of the Page scheme correspond to the serogroups of the Kume scheme). One MAb (E5C12D10) was raised against a Page serovar A strain and the remaining MAbs (F2E6, D6D8D5, and B3E6F9) against a Page serovar C strain. Six different reaction patterns were found among the 76 isolates of H. paragallinarum. There was total correlation between the MAb reaction pattern and the Page scheme, and thus the Kume scheme, to the serogroup level. All 19 Page serovar A (= Kume serogroup A) strains reacted only with MAb E5C12D10, whereas all five Page serovar B (= Kume serogroup B) strains failed to react with any of the MAbs. All 52 remaining strains were Page serovar C (= Kume serogroup C), and all failed to react with MAb E5C12D10 but showed varying reaction patterns with the three other MAbs. Although the MAbs recognized four subdivisions within Kume serogroup C, these subdivisions differed from the four Kume C serovars. This panel of MAbs can be used to assign isolates of H. paragallinarum to either Page serovars or Kume serogroups. Although the subdivisions recognized by the MAbs within the Page serovar C strains do not correspond to the Kume serovars, they may be useful in epidemiological applications.     

Evaluation of two monoclonal antibodies for serotyping Haemophilus paragallinarum

Two monoclonal antibodies (MAbs) were evaluated for their ability to serotype 108 isolates of Haemophilus paragallinarum. One MAb (E5C12D10) was raised against a Page serovar A strain and the other (F2E6) against a Page serovar C strain. In both dot blot and hemagglutination-inhibition tests, MAb E5C12D10 recognized the type strains of Page serovar A and Kume serovars A-1, A-2, A-3, and A-4. MAb F2E6 recognized the type strains of Page serovar C and Kume serovars C-1, C-2, and C-3. Neither antibody recognized the type strains of Page serovar B or Kume serovars B-1 and C-4. When evaluated with 97 field isolates in a dot blot test, the MAbs serotyped 81 isolates, which was better than agglutinin typing by the Page scheme (69 isolates serotyped). The field isolates that did not react with the MAbs were either Page serovar B/Kume serovar B-1 (three isolates), Page serovar C/Kume serovar C-4 (12 isolates), or nontypable by either the Page or Kume scheme (one isolate).     

Serological classification of Japanese isolates of Haemophilus paragallinarum using two serovar-specific monoclonal antibodies

A serological classification of 106 Japanese isolates of Haemophilus paragalinarum recovered from 1960 to 1984 was performed by dot-blotting and hemagglutination-inhibition (HI) tests using two serovar-specific monoclonal antibodies (MAbs), E5C12D10 and F2E6. By the dot-blotting test, 49 of the isolates were serovar A and 55 isolates were serovar C, and the two remaining isolates did not react with either MAb. These two nontypable strains had no hemagglutinating activity against chicken erythrocytes and were nonpathogenic to chickens. Although 49 serovar A isolates were serotyped by the HI test, only 23 of the 55 serovar C isolates could be serotyped. The remaining 32 isolates could not be serotyped because no or low hemagglutinating activity could be detected. Our results indicate that H. paragallinarum serovars A and C have both been present in Japan since 1960, with serovar A isolates being dominant before 1970 and serovar C isolates more prevalent than serovar A since 1970.     

Production and evaluation of a panel of monoclonal antibodies against Haemophilus paragallinarum

We report on the production and characterisation of monoclonal antibodies (MAbs) against Haemophilus paragallinarum, the causative agent of infectious coryza. A bank of 8 MAbs were produced by traditional techniques - four against the reference strain for Page serovar A (0083) and four against the reference strain for Page serovar C (Modesto). Seven of the eight MAbs were shown to be IgG(1) with one being nontypable. None of the MAbs had HI activity and none gave any detectable reaction when examined by Western blotting. None of the MAbs gave a positive reaction in the indirect ELISA with any of the eight type strains of Pasteurella species or sub-species. None of our 8 MAbs gave serovar specific reactions when used in an indirect ELISA format. There was a trend for the serovar A MAbs to give a higher titre with serovar A isolates/strains and a similar trend for the serovar C MAbs to give higher titres with the serovar C isolates/strains.     

Immunogenicity of Haemophilus paragallinarum serovar B strains.

Immunogenicity of three Haemophilus paragallinarum serovar B strains was investigated in cross-protection tests using monovalent vaccines prepared from the B strains, as well as one strain each of serovars A and C. A bivalent vaccine composed of the serovar A and C strains also was used. In the studies with the monovalent vaccines, the immunogenicity of serovar B strains was different from that of serovar A and C strains, although only partial serovar B-specific protection with the three strains was observed. Chickens vaccinated with the bivalent vaccine protected against challenge with one serovar B strain, as well as serovar A and C strains, but not against the other two serovar B strains.     

Identification and expression of a gene encoding an epitope that induces hemagglutination inhibition antibody to Avibacterium paragallinarum serovar A

The aims of this study were the identification, cloning, and expression of a genetic region encoding an epitope that induces hemagglutination inhibition (HI) antibody against Avibacterium paragallinarum serovar A and an evaluation of the recombinant protein for immunogenicity in chickens. Although two monoclonal antibodies (MAbs) with HI activity, designated S24-951 and S7-1716-5C, were generated in this study, no reactive proteins with both MAbs were identified by Western blot analysis. A gene fragment of 5157 bp, designated hpa5. 1, was cloned from genomic DNA, and a recombinant protein expressed by hpa5.1, designated HPA5.1, reacted with both MAbs on dot-blot analysis. HPA5.1 showed no hemagglutinating activity, but significantly absorbed HI antibodies in the chicken immune serum. Analysis using a series of deletion mutants prepared from hpa5.1 indicated that a 4.8 kbp gene in hpa5.1 is essential for the expression epitope recognized by MAb S24-951. In addition, chickens immunized once with HPA5.1 showed a high protection rate with sufficient HI antibody titers against challenge exposure with a virulent strain of A. paragallinarum serovar A strain 221. These results show that hpa5. I1 is responsible for the expression of an epitope that induces HI antibody, and HPA5.1 might be a candidate for the development of a new vaccine against avian infectious coryza caused by A. paragallinarum serovar A.     

Serotyping of chlamydial clinical isolates from birds with monoclonal antibodies

Forty-nine avian chlamydial strains, isolated mainly from various regions in France and from different species of birds, were analyzed and tested with a panel of nine monoclonal antibodies (MAbs) by the indirect microimmunofluorescence test (MIF). The MAbs included five serovar-specific MAbs, three MAbs raised against Chlamydia psittaci and Chlamydia pecorum ovine strains, and one genus-specific MAb. Of the 49 isolates, 41 came from parrots or budgerigars; the rest were from pigeons, a canary, a duck, and a dove. Two additional strains were from unknown hosts. Most of these avian strains were successfully serotyped according to their reactions with five serovar-specific MAbs by the MIF test. The serovars of 44 strains were determined: 39 were of serovar A, 3 of serovar B, and 2 of serovar E. The remaining five isolates were unclassified because they did not react with any of five serovar-specific MAbs but did react with genus MAb or the MAbs produced with ovine strains. The five unclassified isolates (two from budgerigars, two from Gabon gray parrots, and one from a duck) indicate that one or more additional serovars of C. psittaci exist in birds. The heterogeneity within each subgroup was evident because the 49 avian isolates gave 10 subgroups when the results of the five serovar-specific MAbs were combined with results from the three MAbs produced with ovine strains. This heterogeneity of the serovar isolates, as shown by the combination of MAbs, could provide strain markers very useful for epidemiologic studies.     

Hemagglutinating activity and immunological properties of Haemophilus paragallinarum field isolates in Japan

Four field isolates (S4, S10, S15, and S17) of Haemophilus paragallinarum were recovered from chickens affected with infectious coryza in widely separated regions of Japan. Their hemagglutinating (HA) activity and immunological properties were compared with those of strain 221 of serovar A/1 and strains Modesto and S1 of serovar C/2. When treated with potassium thiocyanate or hyaluronidase, all the isolates showed HA activity against formaldehyde-fixed chicken erythrocytes but not against fresh chicken erythrocytes. In the hemagglutination-inhibition (HI) test, the isolates cross-reacted with strains Modesto and S1 but not with strain 221. The immunological properties of these isolates, as determined by cross-protection tests, were similar to those of strain S1 and, to a lesser degree, strain Modesto, but not to strain 221. Our results indicated that the four field isolates belong to serovar C/2 and that the HI test is a suitable method for serotyping H. paragallinarum.