Lymphadenitis In Sheep Research Articles

Caseous Lymphadenitis in sheep and goats – “Cheese Glands”

Caseous lymphadenitis (CLA) is a chronic infectious disease that affects small ruminants and is caused by Corynebacterium pseudotuberculosis. This highly contagious pathogen leads to significant economic losses in the livestock industry due to loss of productivity, rejection of carcasses, and increased veterinary costs. CLA is characterised by the formation of abscesses in lymph nodes and internal organs. Rupture of these nodes can lead to additional contamination of the environment and further transmission. Diagnosis of CLA involves clinical examination, bacterial cultures, serological testing, and advanced molecular techniques for more accurate detection. Treatment options are limited and often ineffective as the pathogen can survive in abscesses and evade the host's immune response. Antibiotic therapy can provide temporary relief but does not eliminate infection, emphasising the importance of prevention measures. Control strategies focus on biosecurity, culling infected animals, and vaccination. While currently available vaccines reduce the incidence and severity of the disease, they do not provide complete immunity and need to be further improved. Understanding the virulence mechanisms of the pathogen and the interactions between the host and pathogen is crucial for the development of more effective vaccines and therapeutic approaches. Ongoing research and new ideas are crucial to reduce the impact of CLA on animal health and the farm economy. This emphasises the need for comprehensive management strategies, including strict hygiene measures, regular checks, and targeted vaccination plans. In addition, due to its zoonotic potential, C. pseudotuberculosis can contaminate meat and milk from infected animals, posing a risk to consumers. The ability of the pathogen to infect both animals and humans emphasises the importance of research into its prevention and diagnosis.

Caseous lymphadenitis in sheep and goats semiotics and the experience of struggle

Based on the analysis of the research experience of various scientists in recent years, this article describes both the characteristic and non-specific pattern of the manifestation of caseous lymphadenitis, possible methods of combating epizootics, including through active immunization of animals. Existing data do not sufficiently disclose the pathogenesis and can be used for preliminary diagnosis, and control methods developed during many years of research and testing do not always effectively solve the problem of epizootics, which is due, among other things, to the lack of vaccines.

Rearing Management and Its Impact on Caseous Lymphadenitis in Sheep.

Two surveys were conducted to assess the prevalence and risk factors of ovine caseous lymphadenitis (CL) and explore the association between its superficial and visceral forms in Sardi sheep in Settat province, Morocco. A total of 1521 sheep, including 318 lambs < 6 months, 572 young sheep aged 6-24 months, and 631 adult sheep > 24 months, were examined from 57 flocks. Superficial CL prevalence levels were as follows: 69/318 in lambs, 292/572 in young sheep, and 114/631 in adult sheep. Additionally, 2124 sheep, including 1813 young and 311 adult sheep, were inspected in slaughterhouses ante- and post-mortem. Among all infected animals, the prevalence of visceral abscesses was notably higher in adult sheep (83%, OR = 9.8, 95% CI = 5.5-17.2) compared to young sheep (35%). Data on flock size, sex, age, body condition score, rearing methods, and hygienic measures were collected. Suspected CL cases were confirmed using quadruplex-PCR. Poor barn ventilation, lack of abscess control, and younger age were identified as key risk factors for CL. Additionally, a high percentage (89%) of animals with thoracic abscesses did not display superficial lesions. While age and abscess control have been reported in previous studies, barn ventilation appears to be a new, but important, risk factor. In conclusion, the identified risk factors in Settat province are linked to breeding management practices. Implementing regular abscess control measures and improving barn ventilation are essential for CL prevention.

Comparative Diagnosis of Caseous Lymphadenitis in Sheep by Molecular and ELISA Method

Bu çalışmada; Diyarbakır yöresindeki koyunlarda Kazeöz Lenfadenitis’in (KLA) etiyolojik ajanı olan Corynebacterium. pseudotuberculosis (C. pseudotuberculosis)’in bakteriyolojik kültür, PCR ve ELISA teknikleri kullanılarak tanısının ortaya konulması, izolasyon bulguları ile testlerin özgüllük ve duyarlılıklarının belirlenmesi ve böylece hastalığın rutin teşhisinde kullanılabilirliklerinin ortaya konulması amaçlanmıştır. Çalışmanın materyalini Diyarbakır’ın farklı bölgelerinde olmak üzere 11 ayrı sürüden toplamda 2650 adet Akkaraman koyun oluşturdu. Sağlık taramasından geçirilen koyunların içinden 90 adet KLA semptomları gösteren ve yüzeysel lenf yumrularından bir veya birkaç tanesi birden apseleşmiş hasta koyunlar tespit edilerek çalışmaya dahil edildi. Kan örnekleri ve apse içeriğinden swap örnekleri alındı. Etkenin tanısı için, swap örneklerinden bakteriyolojik kültür ve PCR analizleri, kan örneklerinden ise ELISA analizleri yapıldı. Diyarbakır yöresindeki koyunlarda PCR analizlerine göre 26 (%28.9) pozitif, 64 (%71.1) negatif, ELISA analizinde 17 (%18.9) pozitif, 73 (%81.1) negatif ve bakteriyolojik kültürde 12 (%13.3)’sinde C. pseudotuberculosis, 16’sından Staphylococcus aureus (S. aureus), 8’inde Streptococ spp. ve 6’sında Escherichia coli (E. coli) kolonileri izole edilirken 48 (%53.3) kültürde hiç üreme olmadığı görüldü. PCR, ELISA ve bakteriyolojik kültür sonuçları karşılaştırıldığında PCR’ın daha belirleyici bir tanı testi olduğu görüldü (p&amp;lt;0.000). İzolasyon sonuçları dikkate alındığında KLA’nın tanısında PCR’ın ELISA’ya oranla duyarlılığı %100, özgüllüğü ise %65 olarak tespit edildi. Sonuç olarak KLA’nın bölge koyunlarında görüldüğü ve bu nedenle bölgede bu hastalığın önlenmesi için kontrol ve eradikasyon çalışmalarının yapılması gerektiği kanaatine varıldı.

Bacteriological, molecular and histopathological evaluation of four vaccines’ protective role against caseous lymphadenitis in sheep

The caseous lymphadenitis disease (CLA) due to Corynebacteriumpseudotuberculosis has worldwide distribution and indicates high prevalence in different countries. The bacterium has many pathogenicity factors; firstly, “phospholipase D”, an exotoxin virulence factor that enhances vascular permeability and facilitates bacterial transmission causing endothelial cell damage. The bacterium's exterior lipid cover is the second pathogenicity factor. This protects it from hydrolytic enzymes found in the phagocytes of the host, where the bacteria proliferate and then release after rupturing. Bacterial proliferation causes CLA abscesses, which are followed by attraction and the formation of an inflammatory response, which increases lymph flow and vascular permeability. The purpose of this study was to look atpostmortem findings, bacterial cultures, and histology of vaccinated sheep to see how well they were protected. On 15 male local sheep bread, four prepared vaccinations against C. pseudotuberculosis biotype 1 were tested. The animals were divided into 5 groups; four of various vaccines used and the fifthwere kept non-vaccinated. A live virulent strain of C. pseudotuberculosis was given to each group. Unvaccinated animals displayed CLA symptoms similar to those seen in spontaneously sick animals.

Bacteriological, molecular and histopathological evaluation of four vaccines’ protective role against caseous lymphadenitis in sheep

The caseous lymphadenitis disease (CLA) due to Corynebacteriumpseudotuberculosis has worldwide distribution and indicates high prevalence in different countries. The bacterium has many pathogenicity factors; firstly, “phospholipase D”, an exotoxin virulence factor that enhances vascular permeability and facilitates bacterial transmission causing endothelial cell damage.The bacterium's exterior lipid cover is the second pathogenicity factor.This protects it from hydrolytic enzymes found in the phagocytes of the host, where the bacteria proliferate and then release after rupturing. Bacterial proliferation causes CLA abscesses, which are followed by attraction and the formation of an inflammatory response, which increases lymph flow and vascular permeability. The purpose of this study was to look atpostmortem findings, bacterial cultures, and histology of vaccinated sheep to see how well they were protected. On 15 male local sheep bread, four prepared vaccinations against C. pseudotuberculosis biotype 1 were tested. The animals were divided into 5 groups; four of various vaccines used and the fifthwere kept non-vaccinated. A live virulent strain of C. pseudotuberculosis was given to each group. Unvaccinated animals displayed CLA symptoms similar to those seen in spontaneously sick animals. Postmortem examination findings, standard bacteriological culture methods were used to isolate C. pseudotuberculosis, as well as tissue preparation for histopathological sections.

Genotyping and genetic diversity of Corynebacterium pseudotuberculosis strains isolated from caseous lymphadenitis in sheep and goat

Genotyping and genetic diversity of Corynebacterium pseudotuberculosis strains isolated from caseous lymphadenitis in sheep and goat

In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent Assay.

Corynebacterium pseudotuberculosis is the etiologic agent of Caseous Lymphadenitis (CLA), a disease leading to severe damage in sheep and goats farming due to the lack of serological diagnosis, treatment, and effective prophylaxis. In this context, several strategies in an attempt to discover new antigens to compose diagnosis assays or vaccines are fundamental. Therefore, this study aimed to use bioinformatics software to evaluate the critical chemical characteristics of unknown proteins of C. pseudotuberculosis by selecting them for heterologous expression in Escherichia coli. For this purpose, six protein sequences of ascorbate transporter subunit, UPF protein, MMPL family transporter, Ribonuclease, Iron ABC transporter domain-containing permease, and fimbrial subunit were obtained. In silico analyses were performed using amino acid sequences to access immunodominant epitopes and their antigenic and allergenic potential and physicochemical characterization. The expressed proteins were used as an antigen for serological diagnosis by ELISA. All proteins showed distinct immunodominant epitopes and potential antigenic characteristics. The only proteins expressed were PTS and Ribonuclease. In parallel, we expressed CP40 and all were used with ELISA antigen in 49 CLA positive sera and 26 CLA negative sera. The proteins alone showed 100% sensitivity and 96.2%, 92.3%, and 88.5% specificity for rPTS, rRibonuclease, and rCP40, respectively. When proteins were combined, they showed 100% sensitivity and 84.6%, 92.3%, 88.5%, and 92.3% specificity for rPTS/rCp40, rRibonuclease/rCP40, rPTS/rRibonuclease, and rPTS/rRibonuclease/rCP40, respectively. The results of this study show an excellent correlation of sensitivity and specificity with all proteins. None of the specificity values preclude the potential of rPTS, rRibonuclease, or rCP40 for use in ELISA diagnostic assays since the results of this work are superior to those of other studies on CLA diagnosis described in the literature.

Virulence Potential, Antimicrobial Susceptibility and Phylogenetic Analysis of Corynebacterium pseudotuberculosis Isolated from Caseous Lymphadenitis in Sheep and Goats in Duhok City, Iraq

Virulence Potential, Antimicrobial Susceptibility and Phylogenetic Analysis of Corynebacterium pseudotuberculosis Isolated from Caseous Lymphadenitis in Sheep and Goats in Duhok City, Iraq

Epidemiological, bacteriological and molecular studies on caseous lymphadenitis in sheep of Dakhlia, Egypt

Caseous lymphadenitis (CLA) is a chronic and insidious disease that mainly affects small ruminants and caused by Corynebacterium pseudotuberculosis (C. pseudotuberculosis). The aims of this research were to identify C. pseudotuberculosis by PCR from pyogenic lesions, to study the phylogenetic analysis of C. pseudotuberculosis and to detect the prevalence based on the detected superficial lesions of CLA in Dakahlia governorate, Egypt. Out of 3471 clinically examined animals, 129 (3.71%) animals were affected with CLA. The isolation rate of C. pseudotuberculosis in abscess of sheep was 45.74% (59/129). Out of 129 samples examined by PCR assay, 63 (48.83%) were positive phospholipase D (PLD) indicated at fragment size 203 bp. This is the first phylogenetic analysis study of C. pseudotuberculosis isolate in Egypt which was isolated from infected sheep. Nucleotide sequence identity data demonstrated that C. pseudotuberculosis PLD gene (MW187942) Dakahlia share homology 99.01%, 98.83 and 98.48% with Zagazig, Egypt (MN867024), Tamil nadu, India (MG720636) and Sudan (MG692441), respectively. In conclusion, this study provided information on the molecular detection and phylogeny of C. pseudotuberculosis in Egypt. Findings of this study can be conducted in other CLA endemic countries with similar animal breeding practices in the Middle East and developing countries.

Risk factors associated with the seroprevalence of caseous lymphadenitis in sheep

This study aimed to determine the prevalence of caseous lymphadenitis (CLA) caused by Corynebacterium pseudotuberculosis (C. Pseudotuberculosis) and its risk factors of sheep reared in four districts (Sherbin, Mansoura, Belkas, and Dekernes) in Dakahlia Governorate. A total of 346 sheep serum samples collected from the study area were examined against antibodies for C. Pseudotuberculosis infection using enzyme-linked immunosorbent assay (ELISA). The prevalence of C. Pseudotuberculosis infection was estimated at 73.98%. The highest prevalence was found among flocks reared in Sherbin (83.65%) followed by those in Mansoura (74.41%) and Belkas (65.47%). A multivariate logistic regression was used to identify risk factor associated with CLA, and the results showed a significant association between the infection and the raising of sheep in open flock not in a closed farms (P = 0.033; odds ratio “OR”: 1.84; 95% confidence interval “CI”: 1.05–3.88). Additionally, higher prevalence was found in herds that keep diseased animals (P = 0.047; OR: 1.98; 95% CI: 1.01–3.23) than those herds which discard diseased animals. The infection by C. Pseudotuberculosis was increased in old age animals than young age ones. On the contrary, gender, treatment, and contact with goats were not associated with the higher prevalence of CLA in sheep (P > 0.05). In summary, the present study determined a high seroprevalence of C. Pseudotuberculosis infection among sheep in Egypt. Farmers should avoid potential risk factors associated with CLA that include keeping diseased animals for breeding and prevent introduction of newly purchased animals from flock of unknown status of infection especially old animals and this may be useful to establish accurate preventive measures.

Factors associated with the seroprevalence of caseous lymphadenitis in sheep from Northeastern Brazil

Caseous lymphadenitis (CL) is one of the main infectious diseases in sheep, responsible for great economic losses, mainly in Northeast region of Brazil, which has the largest sheep flock in the country. Thus, the aim of this survey was to determine the factors associated with the seroprevalence of this disease in sheep from Northeastern Brazil using a planned sample of flocks and animals. Samples were collected from 2312 adult and young sheep of both sex from 196 farms, located in 51 municipalities in five Northeastern Brazilian states (Ceará, Paraíba, Piauí, Rio Grande do Norte, and Sergipe). Serological diagnosis was performed using the indirect ELISA technique. Factors associated with seropositivity were identified by variables extracted from epidemiological questionnaires administered to breeders, using univariable and multivariable analyses. Factors associated with prevalence were purebred sheep (Prevalence ratio - PR = 1.189; P = 0.017), ram acquisition from animal expositions (PR = 1.192; P = 0.020), offspring is separated from ewes (PR = 1.132; P = 0.048), water supplied to sheep from ponds (PR = 1.365; P = 0.002), and delayed disposal of infected animals (PR = 1.263; P = 0.027). Flock-level and animal-level prevalences were 93.88 % (95 % CI = 89.60–96.46%) and 37.46 % (95 % CI = 35.51–39.45%), respectively. In view of the high prevalence found here and based on the associated factors, the implementation of an efficient CL control programme in Northeastern Brazil is recommended.

Update on caseous lymphadenitis in sheep

Background: Caseous lymphadenitis (CLA), caused by the gram-positive bacteria Corynebacterium pseudotuberculosis, has been present in Great Britain since the 1980s and is now considered endemic. CLA is considered to be...

Seroepidemiological study of Caseous lymphadenitis in sheep from the Northeast region of Brazil using an indirect ELISA

The objective of this study was to determine the frequency of seropositivity Corynebacterium pseudotuberculosis in sheep in five states of northeastern Brazil, using an indirect enzyme-linked immunosorbent assay (i-ELISA). Young and adult sheep of both sexes were used. Blood samples were collected from 2638 sheep from 223 herds across all states. For the i-ELISA, antigens produced from the strain of C. pseudotuberculosis BRM 029971, a bacterial isolate from the Northeast region of Brazil, were used. Sensitivity and specificity indexes were calculated for the validation of the test, using as reference 49 and 134 serum samples from sheep known to be positive and negative, respectively. The i-ELISA presented four false-negative and four false-positive results, showing a specificity of 97.01%, a sensitivity of 91.84%, and an accuracy of 95.63%. These results were calculated based on an optical density (OD) cutoff point = 0.138. Of the 2638 sheep tested, 996 (37.76%, 95% CI = 35.93-39.62%) were seropositive, and of the 223 evaluated herds, 210 (94.17%, 95% CI = 90.28-96.56%) seropositive. The i-ELISA showed adequate sensitivity and specificity, proving to be a useful tool in the diagnosis of caseous lymphadenitis (CLA) in sheep. Infection by C. pseudotuberculosis, determined by serology, is disseminated in the sheep herds in the states of Northeast Brazil. Thus, there is a need to implement effective control measures that prevent the spread of infectious agents.

Brazilian Green Propolis as a Therapeutic Agent for the Post-surgical Treatment of Caseous Lymphadenitis in Sheep.

As antibiotics are ineffective when used to treat caseous lymphadenitis, the surgical excision of lesions is often required. Iodine solution (10%) is currently the choice for the post-surgical treatment; however, it may cause histotoxicity. Propolis are resinous substances composed by a mixture of different plants parts and molecules secreted by bees. As green propolis has already proven to possess anti-bacterial and wound healing properties, this study aimed to evaluate the use of a green propolis-based ointment as a therapeutic agent for the post-surgical treatment of caseous lymphadenitis. The caseous lesions of 28 sheep were surgically excised before dividing animals into two groups: (1) iodine-treated animals and (2) sheep treated with an ointment made with a previously characterized green propolis extract. Clinical data of animals, size of the scar area, the presence of moisture and secretion in the surgical wound, the humoral immune response against the bacterium and the susceptibility of C. pseudotuberculosis clinical isolates to the green propolis extract were analyzed. The green propolis-treated group presented complete healing of the surgical wound 1 week before the iodine-treated group. Additionally, animals treated with the green propolis ointment had fewer cases of wound secretion, but it was not statistically different from the iodine-treated group. No clinical signs indicating green propolis toxicity or other side effects were found, associated with a faster and more organized hair recovery by propolis use. The green propolis extract was able to inhibit the growth of 23 from the 27 C. pseudotuberculosis clinical isolates, with minimum inhibitory and minimum bactericide concentrations ranging from 01 to 08 mg/mL, and did not interfere with the humoral immune response against the bacterium. In addition, green propolis was able to inhibit biofilm formation by four of the C. pseudotuberculosis clinical isolates. We concluded that green propolis is a promising therapeutic agent to be used in the post-surgical treatment of caseous lymphadenitis in small ruminants due to its effects on surgical wound healing, hair recovery, inhibition of wound contamination and bacterial growth.

The combination of Corynebacterium pseudotuberculosis recombinant proteins rPLD, rCP01850 and rCP09720 for improved detection of caseous lymphadenitis in sheep by ELISA.

Corynebacterium pseudotuberculosis is the causative agent of caseous lymphadenitis (CLA), a chronic disease of sheep and goats. Current methods for CLA diagnosis cannot identify all infected animals; therefore, the development of an improved diagnosis is essential. We evaluated recombinant phospholipase D (rPLD) protein individually or combined with rCP01850 or rCP09720 proteins for the detection of CLA in sheep. A total of 40 positive and 25 negative sera samples were analysed by ELISA using the recombinant proteins. ELISA using rPLD (E1), rPLD+rCP01850 (E2) and rPLD+rCP09720 (E3) showed 90, 92.5 and 97.5 % sensitivity and 92, 72 and 92 % specificity, respectively. The area under the receiver operating characteristic curves for E1, E2 and E3 was 0.925, 0.882 and 0.990, respectively. ELISA using rPLD +rCP09720 demonstrated the best sensitivity and specificity. Thus, the combination of these recombinant proteins in indirect ELISA has the potential for the diagnosis of CLA in sheep.

New sequence type ST3756 of Staphylococcus aureus subspecies anaerobius as the causative agent of abscessing lymphadenitis in sheep.

Morel's disease is a form of abscessing lymphadenitis of sheep and goats caused by Staphylococcus aureus subspecies anaerobius. In Europe and Africa, the disease is linked to S. aureus of multilocus sequence type 1464. In an outbreak recorded in 2015 in a flock of 530 animals in the district of Nymburk, Czech Republic, Europe, the causative agent was cultured and subsequently confirmed by Maldi-TOF. Neither antibiotic therapy nor surgical interventions met any success, although the strain isolated was found to be sensitive to antibiotics used. Vaccination and revaccination with inactivated autogenous vaccine administered subcutaneously was relatively successful. Subsequent multilocus sequence typing revealed the presence of new S. aureus sequence type 3756, different from 1464 in three out of seven genes typed. The isolate thus represents a new sequence type of Staphylococcus aureus ssp. anaerobius which should be considered as a causative agent of Morel's disease.

Evaluation of specific and non-specific immune response of four vaccines for caseous lymphadenitis in sheep challenged.

Background:Caseous lymphadenitis (CLA) is a serious disease affects sheep and goat, caused by Corynebacterium pseudotuberculosis. Due to it is non-treatable disease, so the effective preventive vaccines are considered a significant way to combat the disease. All strains of C. pseudotuberculosis have several virulence factors that associated with their cell invasion, survival, and proliferation such as phospholipase D (PLD), outer lipid coat, and secreted proteases.Aim:The present study was directed to perform a comparative innate and acquired immune response assessment of different four vaccine formulas to evoke protection against induced (CLA) challenge in sheep.Materials and Methods:Negative ELISA (free of CLA) 15 local breed male (Balady) sheep were divided into five groups, each has received a different vaccine while the control has received saline buffer. The first vaccine composed of toxoid PLD alone the second composed of toxoid PLD with bacterin (formalinkilled bacteria), the third vaccine composed of toxoid PLD plus covaccine 8, while the fourth one composed of toxoid PLD plus locally produced polyvalent clostridial vaccine. The specific immune response was evaluated through lymphocyte proliferation assay using ELISA BrdU kit, while the non-specific response was estimated by superoxide anion production and lysozyme activity assays.Results:The study revealed that PLD toxoid could evoke the highest specific immune response, showing a stimulation index (9.12%). On the other hand, combined toxoid ↱PLD with bacterin followed by PLD toxoid showed a significant increase in the non-specific innate immune response.Conclusion:The present study indicated that the toxoid PLD alone vaccine was most efficient and provided innate and acquired immune response in animals against CLA.

Molecular identification of Corynebacterium pseudotuberculosis in sheep

Caseous lymphadenitis is still a serious zoonotic problem in Turkey. Sheep suffer from the disease with yield loss in wool and meat production. Moreover, with inexperienced laboratory staff, biochemical identification may go unrevealed. The scope of this study was to demonstrate the presence ofCorynebacterium pseudotuberculosisin sheep by PCR. The sampling was conducted via collecting lymph fluids from the lymph node internal pouch wall of 100 sheep that were examined for the presence ofCorynebacterium pseudotuberculosis. Molecular identification of theCorynebacterium pseudotuberculosisisolates was carried out by establishing the presence of the proline iminopeptidase gene. All isolates were confirmed to beCorynebacterium pseudotuberculosisby polymerase chain reaction. The polymerase chain reaction procedure conducted in this research was observed to be reliable and fast, and could be utilized for confirmation of caseous lymphadenitis in sheep as an optional technique to time-consuming biochemical identification methods.

MOLECULAR DIAGNOSIS OF CASEOUS LYMPHADENITIS IN SHEEP AT DAKAHLIA GOVERNORATE, EGYPT

The aim of the present study was to diagnose caseous lymphadenitis in sheep by PCR assay . A total of 1257 sheep from different flocks in Dakahlia governorate were clinically examined during the period of January 2014 to February 2015. sheep were classified according to the status of infection in the flocks to infected , incontact and free ones. 37 needle biopsy samples were randomly collected from 26 diseased, 7 incontact and 4 free examined sheep for PCR assay. rpoB gene was detected in 37 samples (100%) while PLD gene was in 18 samples (48.64%), from which 16 from diseased sheep and two from incontact ones, while not detected in free sheep. The PCR assay was rapid, specific and significant technique as bacterial culture in detecting corynbacterium pseudotuberculosis in needle biopsy samples from sheep