The usefulness of immunohistochemistry to differentiate between nasal carcinoma and lymphoma in cats: 140 cases (1986–2000)

A retrospective evaluation of 232 feline nasal biopsies initially diagnosed as either carcinoma or lymphoma was performed by two pathologists. One or both pathologists disagreed with the original diagnosis in 15 cases (7%), 14 of which had original diagnoses of carcinoma. Out of the 232 cases, 140, including the disputed ones, were subjected to immunohistochemical staining with epithelial and lymphoid markers. Immunohistochemical staining of the 15 disputed cases showed that the original diagnoses were incorrect in 67% (10/15), unverified in 13% (2/15) and correct in 20% (3/15). Among the consensual diagnoses, immunohistochemistry revealed that 3% (4/125) of diagnoses were unverified because they did not stain for any of the markers evaluated. This report demonstrates the importance of immunohistochemistry in establishing a correct histologic diagnosis for nasal neoplasms in cats.     

Gallid-1 herpesvirus infection in the chicken. 3. Reinvestigation of the pathogenesis of infectious laryngotracheitis in acute and early post-acute respiratory disease

Specific-pathogen-free chickens were infected via the trachea when 4 weeks old with 2000 plaque-forming units (PFU) of the virulent Australian infectious laryngotracheitis (ILT) virus strain CSW-1. Titers of ILT virus in the trachea were greatest (10(7.0) PFU/ml in washings, 10(6.0) PFU/g of tissue) 2-4 days postinfection (PI). Infectivity then declined rapidly, to become undetectable by 7 days PI, although highly localized areas of ILT antigen in the tracheal epithelium were occasionally observed by fluorescent antibody staining at 7 and 8 days PI. Tracheal organ cultures established 7 and 8 days PI provided no evidence of latent ILT virus infection at this immediate post-acute stage of pathogenesis. ILT virus was not isolated from peripheral blood leukocytes or lymphoid organs (spleen, bursa, thymus). ILT virus was found in the trigeminal ganglia and/or brain in 14 of 36 chickens (40%) examined between 4 and 7 days after intratracheal inoculation, but it was not in these tissues in five chickens examined at 8 days PI. Virus was also detected at 6 days PI in the trigeminal ganglia in one of five chickens infected by the conjunctival route. These data indicate that the early pathogenesis of ILT (CSW-1) infection frequently involves the tissues of the nervous system. In acute ILT in 4-week-old chickens, interferon-alpha/beta activity was not detectable in serum or tracheal exudates within 14 days PI, but tracheal washings contained significant virus-neutralizing activity by 7 and 8 days PI. In 3-day-old chickens infected via the trachea with 200 PFU of ILT CSW-1, the clearance of ILT virus from the trachea was similar to that observed in 4-week-old chickens, but ILT virus spread systemically to the livers of 20% by 5-7 days PI.     

Histologic incidence and distribution of Cryptosporidium sp. infection in chickens: 68 cases in 1986

In 1986, a retrospective examination of histologic data determined that 68 histology accessions at the Georgia Poultry Laboratories had a diagnosis of cryptosporidiosis. These cases represented 6.4% of the 1065 histologic examinations of chickens performed at the facility. Of the protozoan infections, only Eimeria sp. was diagnosed more frequently (105/1065 = 9.9%). Infected chickens came from 11 companies. Most of the cases were significantly (chi 2 = 103, df = 10, P less than 0.001) associated with two of these companies. These chickens ranged in age from 14 to 51 days. The number of cases per age group was significantly correlated (r = 0.85, t = 3.98, df = 6, P less than 0.01) with the number of cases per age group of the population of histology accessions. Additionally, cases of cryptosporidiosis had a significantly (P less than 0.05) uneven temporal distribution in that few cases were diagnosed during the winter.     

Labeled avidin-biotin enzyme-linked immunosorbent assay for detecting antibody to infectious laryngotracheitis virus in chickens

A labeled avidin-biotin enzyme-linked immunosorbent assay (LAB-ELISA) for detecting antibody to infectious laryngotracheitis (ILT) virus in chicken sera was developed and compared with ordinary ELISA. Purified ILT virus, biotin-labeled anti-chicken IgG rabbit IgG conjugate, and horseradish-peroxidase-labeled avidin were used in the LAB-ELISA. When sera from farm chickens were tested by serum neutralization (SN) and two kinds of ELISA, the correlation rate between SN and LAB-ELISA was 50/50 (100%), and that between SN and ordinary ELISA was 39/50 (78%). In LAB-ELISA, all of the sera that were antibody-negative by SN had low absorbance (A) values (below 0.05), and the A values were closely correlated with the SN indexes. In ordinary ELISA, however, the sera antibody-negative by SN had various A values ranging from 0.06 to 0.32. LAB-ELISA had much lower nonspecific reactions than ordinary ELISA against sera from ILT-negative chickens, even when chickens were 30 weeks old. ILT antibody production after ILT vaccination could be detected by LAB-ELISA. A values peaked 5 weeks postinoculation and were maintained for 17 weeks.     

Correlation of ultrasonographic appearance of lesions and cytologic and histologic diagnoses in splenic aspirates from dogs and cats: 32 cases (2002-2005)

OBJECTIVE: To determine the accuracy of cytologic diagnosis, compared with histologic diagnosis, in determination of disease in ultrasound-guided fine-needle aspirates of splenic lesions. DESIGN: Retrospective study. SAMPLE POPULATION: Splenic specimens from 29 dogs and 3 cats. PROCEDURES: Records were searched for dogs and cats that had undergone ultrasound-guided splenic aspiration. Criteria for inclusion were ultrasonographic identification of splenic lesions and cytologic and histologic evaluation of tissue from the same lesion. Cytologic samples were obtained by fine-needle aspiration, and histologic specimens were obtained via surgical biopsy, ultrasound-guided biopsy, or necropsy. RESULTS: Cytologic diagnoses corresponded with histologic diagnoses in 19 of 31 (61.3%) cases and differed in 5 of 31(16.1%) cases, and 1 aspirate was inadequate for evaluation. In 7 of 31 (22.6%) cases, histologic evaluation of tissue architecture was required to distinguish between reactive and neoplastic conditions. On the basis of histologic diagnosis in 14 animals with nonneoplastic conditions, the cytologic diagnosis was correct in 11 cases, not definitive in 2 cases, and incorrect in 1 case. In 17 animals with malignant neoplastic diseases, the cytologic diagnosis was correct in 8 cases, not definitive but consistent with possible neoplasia in 5 cases, and incorrect in 4 cases. Multiple similar-appearing nodules were significantly associated with malignancy, whereas single lesions were more often benign. CONCLUSIONS AND CLINICAL RELEVANCE: Ultrasound-guided aspiration of splenic lesions is a minimally invasive tool for obtaining specimens for cytologic evaluation. Although cytologic diagnoses often reflect histologic results, if missampling or incomplete sampling occurs or tissue architecture is required to distinguish between reactive and neoplastic conditions, accurate diagnosis with fine-needle aspiration may not be possible.     

Safety and efficacy of the cell-associated vaccine prepared by an attenuated infectious laryngotracheitis virus.

The safety and efficacy of the cell-associated (C-A) vaccine prepared by chicken embryo fibroblast (CEF) cells infected with the tissue-culture-modified strain of infectious laryngotracheitis (ILT) virus were studied in chickens. Over seventy percent of chickens inoculated with the C-A vaccine by the subcutaneous (S.C.) or intramuscular (I.M.) route at 1 day of age was protected against challenge with a virulent strain of ILT virus without any clinical signs. Chickens vaccinated with the C-A vaccine at 1 day of age acquired immunity within 6 days after vaccination, and the protection rate maintained more than 60% until 10 weeks post-vaccination. The C-A vaccine was invariably effective for chickens at various age. There was no evidence that the development of immunity was hindered by further vaccination with Newcastle disease and infectious bronchitis combined live vaccine. In addition, the C-A vaccine was safe when chickens were inoculated with 10 doses. In the field trials of the C-A vaccine, no adverse reaction was observed, and over 65% of vaccinated chickens was protected against the challenge of the virulent ILT virus at 8 weeks after vaccination.     

Studies on the live-virus vaccine against infectious laryngotracheitis of chickens. II. Evaluation of the tissue-culture-modified strain C7 in laboratory and field trials

Laboratory and field tests were conducted to evaluate the immunogenicity of a live-virus vaccine against infectious laryngotracheitis (ILT) of chickens that was prepared using tissue-culture-modified strain C7. Eighty-three percent or more of chickens vaccinated by the ocular (OC) or intranasal (IN) route with 10(4.75) TCID50 of the attenuated strain C7 at 50 days of age were protected against challenge with a virulent strain of ILT virus without showing any clinical signs for 4 weeks post-vaccination (PV). When vaccine was administered by aerosol, however, only 65% of vaccinated chickens were protected against challenge. Fifty-seven percent of chickens vaccinated at 70 days of age maintained immunity for 6 months PV. Immune response in younger chickens was inferior to that in older ones. In the field trials, clinical observation revealed no adverse reactions caused by the vaccination, and 60% or more of broilers and 80% or more of layers vaccinated by the OC route were protected against challenge at 4 weeks PV. These results confirmed the safety and efficacy of the vaccine.     

抗禽流感或传染性喉气管炎两种重组鸡痘病毒疫苗联合免疫的相互影响

“表达H5N1禽流感病毒HA和NA基因的重组鸡痘病毒（rFPV—AI）疫苗”以及“表达传染性喉气管炎病毒gB基因的重组鸡痘病毒（rFPV—ILT）疫苗”均已在实现商业化生产。由于两种疫苗使用了相同的载体病毒,如何使用才能减少相互干扰而产生最好的免疫效果。本研究设计了三个试验组：1）免疫rFPV—AI后间隔4周接种rFPV—ILT;2）rFPV—AI和rFPV—ILT混合后接种;3）将rFPV—AI和rFPV—ILT分别在两个翅膀同时免疫。结果表明,试验鸡接种rFPV—AI后4周接种rFPV—ILT,对传染性喉气管炎病毒WG株攻击的保护率为60％（6／10）,低于rFPV—ILT单独免疫组的保护率（100％,10／10）;rFPV—AI和rFPV—ILT混合后免疫组对禽流感病毒强毒攻击的保护率为80％（8／10）,对传染性喉气管炎病毒强毒攻击的保护率为70％（7／10）,而rFPV—AI和rFPV—ILT分两点同时接种对两种病毒攻击均能产生完全保护。本试验结果建议将这两种相同载体的重组病毒疫苗分两点同时接种以避免相互之间的干扰。     

Detection of infectious laryngotracheitis virus in formalin-fixed, paraffin-embedded tissues by nested polymerase chain reaction

Infectious laryngotracheitis virus (ILTV) is routinely diagnosed by histopathologic examination of trachea, eyelid, and lung tissues. Lesions consistent with infectious laryngotracheitis (ILT) infection include syncytial cell formation with intranuclear inclusion bodies. These changes are present during the acute phase of infection. To increase the sensitivity of detecting ILT, a nested polymerase chain reaction (PCR) was developed for detection of ILTV DNA. Nested PCR assay was specific for the amplification of ILTV DNA and did not amplify a variety of other avian pathogens. To further validate the ability of this assay to detect ILT, nested PCR was performed in formalin-fixed, paraffin-embedded tissues from 35 cases of respiratory disease. Of the 35 cases, 12 were considered ILT suspects on the basis of initial clinical observation. Eleven of the 12 ILT-suspect cases were diagnosed as ILT, and the remaining 24 were diagnosed as nonspecific tracheitis (NST) by histopathologic examination. Histopathologically positive samples were confirmed by direct fluorescent antibody test and virus isolation. Of the 11 ILT-positive cases, 10 were positive by nested PCR. In addition, ILTV DNA was detected in 7 of the 24 samples diagnosed as NST upon histopathologic examination. Therefore, by nested PCR, ILTV DNA was detected in tissues independently of the presence of syncytial cells, intranuclear inclusions, or both. ILT nested PCR is a specific and sensitive assay capable of detecting ILT at different stages of infection and can be utilized in combination with histopathological examination to accelerate the diagnosis of ILT infection.     

Comparison of diagnostics techniques in an outbreak of infectious laryngotracheitis from meat chickens

Various diagnostics techniques were compared for their ability to detect infectious laryngotracheitis (ILT) during an outbreak in chickens aged between 4 and 21 wk. Gross lesions ranged from excess mucus to accumulation of fibrinonecrotic exudate in the larynx and trachea. Syncytial cells with intranuclear inclusion bodies were found in sinus, conjunctiva, larynx, trachea, lung, and air sac. Virus isolation in chicken embryos was attempted in every case. Negative-stain electron microscopy detected herpesvirus in only 6% of the cases. Yet, isolation of ILT virus in the chorioallantoic membrane was presumed by histology in >20% of the samples and confirmed by fluorescent antibody (FA) in 35% of the embryos inoculated with conjunctivas or tracheas from affected birds. Overall, results from histology and FA tests were highly correlated. FA test has the advantage over histology of being diagnostically specific for ILT virus. Polymerase chain reaction was the most sensitive test and detected the viral DNA even in cases where histology and FA were negative. ILT virus DNA was quantified by real-time polymerase chain reaction (Re-Ti ILTV). Histologic and FA results from larynx and trachea were negative if the concentration of the viral DNA was < or =4 of log10. A viral DNA concentration higher than log10 4, as determined by Re-Ti ILTV, was required for clinical ILT to be manifested.     

鸡骨型白血病免疫琼扩与放射摄片诊断的比较

为了探索特异性免疫学诊断方法对鸡骨型白血病(OP)的检测效果。用双盲法分批采样同时进行羽髓琼脂扩散试验和胫骨X线摄片检查对比,共检834只鸡(433羽属天然自发OP鸡群,401羽属人工诱发实验鸡群)。结果羽髓琼扩试验的OP阳性检出率为3．48％(29／834：)。阳性病理符合率为75．86％(22／29)。阳性误诊(假阳性)率24．13％(7／29),阴性误诊(假阴性)率36．15％(291／805)。而X线摄片检查的OP阳性检出率为38．37％(320／834)。X线阳性完全可获病理组织学证实,不存在假阳性或假阴性。把禽白血病羽髓琼扩试验试用于检测鸡骨型白血病尚属初次报道,其阳性检出率低而误诊率高,尚不宜作OP检测手段。放射摄片诊断的阳性检出率高而误诊率低,可快速诊断,故适用于OP检疫。     

The relationship of Cryptosporidium sp. infection of the bursa of Fabricius, intestinal tract, and respiratory system of chickens in Georgia, 1974-1988

During 1974-1984, cryptosporidiosis was diagnosed in slightly more than 1% (63/6050 = 1.04%) of histopathology reports on chickens and turkeys made at two Georgia diagnostic laboratories. During 1985-1988, cryptosporidiosis was diagnosed in almost 6% (157/2622 = 5.99%) of histopathology reports on chickens and turkeys made at the two laboratories. Intestinal and respiratory Cryptosporidium sp. infections have increased significantly (p less than 0.01) with time. During the same period, there was no statistical pattern for bursa of Fabricius Cryptosporidium sp. infections, and there was no relationship between the incidence of bursal infections and the incidence of either respiratory or intestinal infections.     

SPRAY VACCINATION OF CHICKENS USING INFECTIOUS LARYNGOTRACHEITIS VIRUS

Day-old chickens were vaccinated with the SA2 strain of infectious laryngotracheitis vaccine using a coarse spray and 14-day-old chickens were vaccinated using a coarse or a fine (Turbair Vaccinair 240) spray. The Conjunctiva was the most common site of infection in all cases but this almost invariably led to involvement of the nasolacrimal ducts. Other sites in the nasal cavity were affected less frequently. The clinical consequences of infection in day-old chickens were too severe for field use but the mortality (0 to 1.4% in different experiments) in 14-day-old chickens may be acceptable in some circumstances although about 9% developed infection of the trachea. It is concluded that a less pathogenic vaccine is needed if spray vaccination with ILT is to be recommended as a routine procedure particularly for day-old chickens.     

Diagnostic value of cytologic examination of gastrointestinal tract tumors in dogs and cats: 83 cases (2001-2004)

OBJECTIVE: To determine results of cytologic examination of fine-needle aspirates and impression smears of gastrointestinal tract tumors in dogs and cats. DESIGN: Retrospective case series. ANIMALS: 38 dogs and 44 cats with histologically confirmed gastrointestinal tract tumors. PROCEDURES: Results of cytologic examination of fine-needle aspirates (n = 67) or impression smears (31) were compared with the histologic diagnosis, and extent of agreement was classified as complete, partial, none, or undetermined. RESULTS: For 48 of the 67 (72%) fine-needle aspirates, there was complete or partial agreement between the cytologic and histologic diagnoses. For 12 (18%) aspirates, the extent of agreement could not be determined because the cytologic specimen was considered unsatisfactory. For 29 of the 31 (94%) impression smears, there was complete agreement between the cytologic and histologic diagnoses, and for 2 (6%), there was partial agreement. None of the impression smears were considered unsatisfactory. Proportion of samples with complete agreement and proportion of samples with complete or partial agreement were significantly higher for impression smears than for fine-needle aspirates. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that there was moderate agreement between results of cytologic examination of fine-needle aspirates from dogs and cats with gastrointestinal tract neoplasia and the definitive histologic diagnosis. The agreement between results of cytologic examination of impression smears and the histologic diagnosis appeared to be higher.     

Development and evaluation of an ELISA for the detection of antibody to infectious laryngotracheitis virus in chickens

An enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to infectious laryngotracheitis (ILT) virus in chickens was developed and compared with the serum-neutralization assay. The ELISA routinely yielded 16-to-32-fold higher titers than the serum-neutralization test. To overcome the requirement for large amounts of purified viral antigen, the microtiter trays were initially coated with an antibody prepared against purified ILT virus. A relatively crude viral preparation could then be used to coat the trays. Sera from specific-pathogen-free chickens less than 12 weeks of age did not show nonspecific binding, although 2.7% of all sera from chickens between 13 and 64 weeks of age had nonspecific activity. The majority of nonspecific reactors came from one highly inbred flock of specific-pathogen-free chickens. A number of modifications of ELISA procedures reported to reduce the nonspecific binding of chicken sera were investigated. Treatment of the serum or the plate and changes in the composition of the diluent did not increase the relative sensitivity of the anti-ILT assay.     

Increased virulence of modified-live infectious laryngotracheitis vaccine virus following bird-to-bird passage.

Modified-live (ML) infectious laryngotracheitis (ILT) vaccine viruses, both tissue-culture-origin (TCO) and chicken-embryo-origin (CEO), were passaged 20 times in specific-pathogen-free chickens. After serial bird-to-bird passage, increased virulence was observed for CEO virus but not TCO virus. Increased mortality and increased severity and duration of respiratory disease were observed in chickens inoculated with chicken-passaged CEO viruses; only mild respiratory disease (no mortality) occurred in chickens inoculated with chicken-passaged TCO viruses. These findings suggest that ML ILT vaccine viruses may increase in virulence after bird-to-bird passage.     

鸡传染性喉气管炎病毒(江苏株)的分离与初步鉴定

本研究从江苏省某养鸡场采集临床疑似鸡传染性喉气管炎(infectious laryngotracheitis,ILT)的病鸡喉头气管,接种鸡胚绒毛尿囊膜分离病毒,命名为LJS091151。以分离毒人工感染SPF鸡,于攻毒后d3出现了ILT的典型临床症状,剖检亦可见喉头气管黏膜充血肿胀、消化道空虚、黏膜充血等病理变化。根据GenBank发表的鸡传染性喉气管炎病毒(Infectiouslaryngotracheitis virus,ILT)gB、TK基因保守序列设计2对引物对分离株基因组DNA进行扩增和序列测定。序列分析表明,获得的核苷酸序列及其推导的氨基酸序列与GenBank发表的序列相似性均在99%以上,证明了分离株LJS091151为鸡传染性喉气管炎病毒。     

Pathogenicity of infectious laryngotracheitis virus as measured by chicken embryo inoculation

A comparison was made between pathogenicities for chicken embryos of unattenuated and attenuated strains or isolates of infectious laryngotracheitis (ILT) virus. All 11-day-old chicken embryos inoculated with 10(3.0) or 10(4.0) TCID50 of unattenuated strain NS175 via allantoic cavity died within 6 days. On the contrary, no chicken embryos of the same age died when inoculated with the same amount of cell-culture-attenuated isolate C7 in a like manner. The mortality index for chicken embryos (MICE) was obtained by dividing the cumulative number of embryos dying within 7 days by the cumulative number of embryos surviving 7 days. The reliability of the MICE test was confirmed by duplicate and triplicate experiments with strain NS175 and isolate C7. MICE obtained in the experiments with 9 different strains or isolates of ILT virus ranged from 0 to 1, and the values were well correlated with the pathogenicities for chickens. The results from the present work suggest that strains or isolates with MICE less than 0.16 would have low or no pathogenicity for chickens, and those with MICE more than 0.27 would be highly pathogenic. Further studies are needed using additional isolates of ILT virus with varied pathogenicities.     

Studies of infectious laryngotracheitis vaccines: immunity in layers

Ten-week-old layer chickens obtained from a commercial source were eye-drop vaccinated with chicken-embryo-origin (CEO) or tissue-culture-origin (TCO) vaccines for infectious laryngotracheitis (ILT). Controls were not vaccinated. Approximately one-third of the layers were challenged with virulent ILT virus at 21, 40, or 60 weeks of age. Serum samples taken from the layers before challenge were used in a virus neutralization (VN) test to determine vaccination titers at those three ages. Both vaccines induced low VN titers (geometric mean titer [GMT] less than 6). At 21 weeks of age, the titers produced by the two vaccines were not significantly different, but at 40 and 60 weeks of age the VN GMT of the CEO-vaccinated group was significantly greater than that of the TCO-vaccinated group. The VN GMTs did not drop over time in either group and actually rose between 21 and 60 weeks of age in the CEO group. Both vaccines protected layers against severe challenge with virulent ILT virus, neither being significantly better than the other under these experimental conditions. Unvaccinated sentinel chickens were maintained in contact with the vaccinated layers during three intervals between 1 day and 6 weeks post-vaccination. Diagnostic tests performed on the sentinels to detect lateral spread of vaccine virus from vaccinated to unvaccinated chickens showed scattered positive results.     

Retrospective and prospective studies of transmissible viral proventriculitis in broiler chickens in Brazil

We investigated the occurrence and pathologic findings of transmissible viral proventriculitis (TVP) associated with the chicken proventricular necrosis virus (CPNV) in commercial broiler chickens in southeastern Brazil. Seventy-three broilers, 25–36 d old, with a history of reduced growth, were referred to our veterinary pathology services from 2013 to 2017. Broilers were clinically examined, weighed, and euthanized for postmortem examination. Broilers of different ages with proventricular histologic lesions were positive for CPNV by RT-PCR; however, the intensity of histologic lesions was higher among 33-d-old animals, and viral RNA detection was more frequent among those that were 28 d old. In the proventriculi of 35 of 73 (48%) broilers, lesions were characterized by glandular epithelial necrosis, lymphoplasmacytic and histiocytic infiltrates, and metaplasia of glandular epithelium to ductal epithelium. In 24 of 73 (36%) broilers with histologic TVP-compatible lesions, CPNV was detected by RT-PCR for the viral protein 1 (VP1) gene. Broilers with histologic lesions were lighter than expected compared to the Cobb 500 standard weight. TVP has not been reported previously in broiler chickens in Brazil, to our knowledge.