Study on prevalence and bacterial etiology of mastitis, and effects of subclinical mastitis and stage of lactation on SCC in dairy goats in Egypt.

In Egypt, inadequate information on prevalence and epidemiology of caprine mastitis is available. This study was designed to investigate prevalence and etiological agents of caprine mastitis and assess the efficacy of somatic cell count (SCC) as marker of subclinical mastitis (SCM) in dairy goats. This study was carried out on 249 randomly selected lactating goats in different lactation stages and examined clinically. Of these animals, 477 milk samples were aseptically collected and screened for bacterial carriage. SCC was assessed in 234 apparently normal milk samples, and SCC ≥ 106 cells/ml was indicator for SCM. Prevalence of clinical mastitis (CM) was 33.73% and 16.87% at animal and udder-half levels, respectively. SCM was 52.56% in the apparently healthy halves. Culture results proved single infection in 49.69% of samples, mixed infection in 23.9% of samples, and 26.41% of samples were negative. Coagulase negative staphylococci (CNS) were the most predominant bacteria (58.75%), then Staphylococcus aureus (S. aureus) (24.375%), and Streptococci (1.875%) were the least. No significant difference was recorded between mean of SCC in bacteriologically positive and negative samples, neither in those with SCC ≤ 106 nor with SCC ≥ 106 cells/ml both in middle and late lactation stages. Besides, the percentage of animals harboring SCC ≥ 106 cells/ml and negative for bacteriology in late lactation stage was 3 times (28.57%) more than in midlactation (9.3%). We can assume that SCC is not proper indicator for intra-mammary inflammation (IMI) in goats, and bacteriological examination remains more efficient, despites being time consuming and expensive.

Efficacy and toxicity of plasmonic photothermal therapy (PPTT) using gold nanorods (GNRs) against mammary tumors in dogs and cats.

Plasmonic photothermal therapy (PPTT) was introduced as a promising treatment of cancer. This work was conducted to evaluate the cytotoxic effect of intratumoral (IT) injection of 75µg gold nanorods (GNRs)/kg of body weight followed by direct exposure to 2 w/cm2 near infra-red laser light for 10min on ablation of mammary tumor in 10 dogs and 6 cats. Complete blood count (CBC), liver and kidney function were checked before the start of treatment and one month after injection of GNRs. Results showed that 62.5% (10/16), 25% (4/16) and 12.5% (2/16) of treated animals showed complete remission, partial remission and no response, respectively. Tumor was relapsed in 4 cases of initially responding animals (25%). Overall survival rate was extended to 315.5±20.5days. GNRs have no toxic effect on blood profile, liver or kidney functions. In conclusion, GNRs can be safely used for treatment of mammary tumors in dogs and cats.

African Swine Fever and Its Control Measures in Wild Boar: A "De Iure Condito" Analysis in the European Union.

Recently, the African swine fever (ASF) epizootic has been reported in domestic pigs and wild boars in several European Union Member States (EU MS) and epidemiological evidence has accumulated which indicates that wild boar play a key role in maintaining and spreading the disease. Thanks to the experience gained when managing ASF outbreaks in Sardinia (Italy) and Eastern Europe, Directive 2002/60 CE was issued. This directive represented an important step forward in controlling the disease, particularly the risk of spreading the virus to wild animals. Since 2021, according to Regulation (EU) 2016/429, which is also called "Animal Health Law-AHL", when the MS competent authority suspects or confirms ASF (a cat. A listed disease) in wild animals, it is mandatory to conduct surveillance in the wild boar population and implement the necessary control measures. Within AHL, Regulations (EU) 2020/687 and 2023/594 established special ASF control measures in kept and wild porcine animals, and their products and by-products, focusing on and underlying old and new responsibilities that vets (both public and private ones) have to accomplish under the new regulations. The new change in the legal framework deals with specific measures to be applied in the wild and represents a great challenge for MS veterinary services. Some of these measures have been well established in the last two decades, particularly those related to application in the farming system, while other measures are still new to veterinary health management and require a holistic approach in terms of intensity, considering all geographical, ecological, productive, cultural and social features of the involved EU MS. In this contribution, the authors intend to focus on specific measures which have been issued in order to limit or stop the spread of ASF in a wild, "boundless" ecosystem. These measures expand the field of competence of the official veterinary service to wild areas in addition to farm activity.

First Molecular Survey to Detect Mycoplasma gallisepticum and Mycoplasma synoviae in Poultry Farms in a Strategic Production District of Sicily (South-Italy).

Mycoplasmas are recognized as avian pathogens, which may cause both respiratory disease and synovial infections in poultry, resulting in severe economic losses. Our study aims to determine the occurrence of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) among commercial and rural laying hens located in Ragusa province (South Italy), using a duplex real time PCR. Four hundred tracheal swabs were collected from seven commercial (200 swabs) and 25 rural (200 swabs) farms without any clinical disease history. Out of 400 swabs collected, 50 (12.5%) and 93 (23.25%) were positive for MG and MS, respectively. In particular, 9 (18%) and 22 (23.65%) positive swabs for MG and MS, respectively, originated from commercial farms, compared to 41 (82%) and 71 (76.34%) obtained from rural farms. Data obtained show a lower prevalence of MG than MS in the studied farms. Moreover, both pathogens were spread in rural and commercial farms. PCR could be concluded as a rapid and sensitive method for the identification of MG and MS in areas where commercial farms that are declared Mycoplasma-free and rural flocks coexist. These data highlight the importance of surveillance also in rural poultry to monitoring the occurrence of mycoplasmas strains in strategic productive districts.

Preliminary Study on the Host Response to Bivalent and Monovalent Autogenous Vaccines against Mycoplasma agalactiae in Dairy Sheep.

In Italy, dairy sheep farming represents a vital agro-industry sector, but it is still challenged by contagious agalactia (CA), which is endemic there, and vaccination is the most economical and sustainable tool for control. This study aimed to evaluate the combined Mycoplasma agalactiae (Ma)-Staphylococcus aureus (Sa) vaccine (Ma-Sa) against the Ma monovalent vaccine in ewes. Twelve primiparous Ma-free ewes were randomly grouped into three equal groups: first, the control group injected with placebo, second, the group vaccinated with the Ma monovalent vaccine, and third, the group vaccinated with Ma-Sa combined vaccine, with two S/C doses at 45-day intervals. The animals were examined for serological, hematological, and somatic cell count (SCC) changes for 17 successive weeks. A significant increase in anti-Ma antibody mean titers, leukocytes, and platelets was observed in the vaccinated animals, with the highest values in those who received the combined vaccine. Neutrophils were high only in the animals who received the combined vaccine. SCC was lower in the vaccinated animals during the first six weeks. This study concludes that the combined Ma-Sa vaccines enhance immune response and potentiate its efficacy against Ma. This improvement might be attributed to the sensitization/activation effect of S. aureus on platelets, which are recoded to act as a key regulator for the coordination of all components of the innate immune system. Even though this study included a small number of animals, its findings about the potentialities of this inactivated vaccine in the control of CA are strongly encouraging. Further confirmation might be needed through additional replicates and a challenge study is needed before proceeding with widespread use.

Diagnostic Findings in a Confirmed Outbreak of Brucella ovis Infection in a Traditional Sheep Farm in Sicily (South-Italy).

Aim of this study is to report a laboratory investigation performed following the isolation of Brucella ovis, causing ovine epididymitis, in a traditional sheep farm in Sicily (South Italy). This disease represents a newly emerging risk for Italian livestock and is listed among diseases of EU priority (EU Reg 2016/429). Blood samples from 56 rams and 143 ewes were analyzed by both Enzyme-Linked Immunosorbent Assay (ELISA) and Complement Fixation Test (CFT). Genital swabs from all rams and 15 lactating ewes were collected to perform real-time PCR. Eighteen serologically positive rams were slaughtered and postmortem-inspected. Samples of testicle, epididymis, lymph nodes, and urine were also collected in order to perform microbiological, molecular, and histopathological analysis. Twelve slaughtered rams showed anatomo-pathological lesions. Real-time PCR for B. ovis BOV_A0504 gene was positive for 13 testicles and epididymis and 11 urine while B. ovis was isolated from epididymis and testicles of 7 slaughtered rams. This is the first exhaustive laboratory report of a microbiological, molecular, and serological pattern of the disease in sheep in Italy. Despite the impact on health and animal welfare, the epidemiology of B. ovis infection is still unknown, particularly in our country where the disease is considered endemic.

Novel Biomarkers of Mastitis in Goat Milk Revealed by MALDI-TOF-MS-Based Peptide Profiling.

Mastitis is the most common infection of dairy goats impairing milk production and quality, which is usually recognized by mammary gland visual inspection and palpation. Subclinical forms of the disease are also widely represented, which lack the typical signs of the clinical ones but are still associated with reduced production and safety for human consumption of milk, generally presenting a high bacterial count. In order to obtain novel analytical tools for rapid and non-invasive diagnosis of mastitis in goats, we analyzed milk samples from healthy, subclinical and clinical mastitic animals with a MALDI-TOF-MS-based peptidomic platform, generating disease group-specific spectral profiles whose signal intensity and mass values were analyzed by statistics. Peculiar spectral signatures of mastitis with respect to the control were identified, while no significant spectral differences were observed between clinical and subclinical milk samples. Discriminant signals were assigned to specific peptides through nanoLC-ESI-Q-Orbitrap-MS/MS experiments. Some of these molecules were predicted to have an antimicrobial activity based on their strong similarity with homolog bioactive compounds from other mammals. Through the definition of a panel of peptide biomarkers, this study provides a very rapid and low-cost method to routinely detect mastitic milk samples even though no evident clinical signs in the mammary gland are observed.

Q fever: A neglected disease of camels in Giza and Cairo Provinces, Egypt.

BACKGROUND AND AIM: Q fever is a zoonotic disease caused by Coxiella burnetii. Cattle, sheep, and goat are the main reservoir of C. burnetii. In Egypt, the epidemiological data about C. burnetii in camels are limited. Therefore, the current study was conducted to identify C. burnetii infection in camels by different molecular tools and to estimate its seropositivity through the detection of anti-C. burnetii antibodies in camel sera. MATERIALS AND METHODS: Blood samples were collected 112 from camels in Giza and Cairo Provinces, Egypt. All blood samples were screened by trans-quantitative polymerase chain reaction (trans-qPCR) for C. burnetii and positive samples subjected to standard PCR using the superoxide dismutase enzyme coding gene of C. burnetii. Sera of studied camels were examined for the presence of antibodies against C. burnetii using enzyme-linked immunosorbent assay. RESULTS: Out of 112 camels, 19 were positive for C. burnetii by qPCR with an overall prevalence of 16.9% (18.6% in Giza and 15.1% in Cairo Provinces, respectively). The seroprevalence of anti-C. burnetii IgG antibodies in the examined camels was 4.5% (5/112). CONCLUSIONS: Trans-qPCR assay is a rapid and sensitive tool for the detection of C. burnetii in acute stage. Camels should be considered one of the major reservoirs for C. burnetii in Egypt.

Corynebacterium pseudotuberculosis mastitis in Egyptian dairy goats.

BACKGROUND AND AIM: Mastitis is an important threat facing goat milk industry and is the most common cause of culling. Efficient control of mastitis, based on efficient diagnosis of diseased animals, would improve milk production and reproductive efficiency. In subclinical mastitis (SCM), infected goats demonstrate neither udder symptoms nor abnormal milk. Corynebacterium pseudotuberculosis is an infectious causative agent of mastitis, mostly results as an extension of infection from the supramammary lymph node, and causes financial losses in the goat industry. This study aimed to estimate the prevalence of SCM with emphasis on C. pseudotuberculosis mastitis in Egyptian dairy goats in the selected farms. MATERIALS AND METHODS: A total of 336 half milk samples were collected from 177 dairy goats of various crossbreeds, in mid-to-late lactation period, after clinical examination. All samples were examined bacteriologically, while somatic cell count (SCC) was determined only in 180 half milk samples of the clinically healthy milk samples. The isolated and identified C. pseudotuberculosis was examined for evidence of virulence genes (Phospholipase D [pld] and ß-subunit of RNA polymerase [rpoB]) by polymerase chain reaction (PCR). RESULTS: The prevalence of clinical mastitis was 30.5%, while 69.5% of animals were apparently healthy and secreted milk was normal. Of those 180 clinically healthy half milk samples, 96 milk samples (53.33%) showed SCM as detected by SCC (SCC ≥1,000,000 cells/ml). Coagulase-negative staphylococci were the most prevalent bacteria (41.96%), then Staphylococcus aureus (37.5%) and C. pseudotuberculosis (7.14%). Molecular diagnosis of virulence genes revealed evidence of pld gene in 16 isolates (66.66%), and rpoB gene in 6 samples (25%) of the 24 bacteriologically isolated C. pseudotuberculosis. Here, we describe, for the 1st time, isolation and identification of C. pseudotuberculosis from milk of does suffering from SCM in Egypt. CONCLUSION: C. pseudotuberculosis must be considered for routine bacteriological examination of milk from dairy goats, particularly herds with a history of caseous lymphadenitis. Pld gene-based PCR is more reliable than rpoB gene-based ones for the diagnosis of C. pseudotuberculosis.

Seroprevalence, isolation, molecular detection and genetic diversity of Toxoplasma gondii from small ruminants in Egypt.

Toxoplasmosis is an infectious zoonotic disease caused by protozoan Toxoplasma gondii. Detection of T. gondii infection with touchy and particular strategies is a key advance to control and prevent toxoplasmosis. Genotyping can explain the virulence, epidemiology and setting up new methodologies for diagnosis and control in human and animals. The point of this study was to assess the seroprevalence of T. gondii in sheep and goat in Egypt and to comprehend the genetic variety of T. gondii isolates circling in Egypt. Blood samples were gathered from 113 ewes and 95 she-goats from three Egyptian governorates (Cairo, Giza and Al-Sharkia). Also blood and tissue samples were gathered from 193 sheep and 51 goats from Cairo and Giza abattoirs. All samples were assayed serologically utilizing ELISA and OnSite Toxo IgG/IgM Rapid test cassettes (OTRT) tests and the tissue samples of the seropositive animals were digested and microscopically examined then bio-assayed in mice as viability test. All the T. gondii isolates undergo molecular identification using PCR and genotyped utilizing nPCR/RFLP analysis of SAG2 gene. The total seropositivity of live sheep and goat was 47.15 and 39.2% utilizing ELISA and OTRT respectively. Concerning abattoirs, seropositivity, positive microscopic examination, mice viability from sheep samples were 47.1%, 37.3% and 44.1% respectively while that of goats were 45.5%, 33.3% and 48.6% respectively. Eighteen T. gondii isolates were affirmed utilizing PCR. Genotyping confirmed 10 isolates (55.5%) as type II, 6 (33.3%) as type III and 2 (11.1%) as atypical genotypes. Type II and III are the genotypes mostly circling among small ruminants in Egypt and this is most significance for the public health in Egypt.