Morphology of the third eyelid and superficial gland of the third eyelid on pig fetuses

The morphological and histological examinations of the third eyelid and superficial gland of the third eyelid were conducted in pig fetuses coming from the 20th, 24th, 27th, 30th, 35th, 50th, 63rd, 94th and 112th day of gestation. The morphological examinations were carried out by applying the method of macroscopic preparation with a forehead magnifying glass and binocular (magnification 1.5-5.0x). In order to make anatomical elements more visible, 60-80% absolute alcohol and 0.5-4% acetic acid solution were used for the examinations. On the 20th, 24th, 27th and 30th day of gestation, the whole fetuses were collected for the histological examinations. The whole eyeball with developing accessory organs was collected from the pig fetuses on the 35th day of gestation. On the 50th, 63rd, 94th and 112th day of gestation, only the superficial gland with the third eyelid was collected. Staining with haematoxylin-eosin (H-E) and Azan method was performed. On the 20th, 24th, 27th and 30th day of gestation, the primordia of the glandular epithelium were not found in the examined material. The process of the third eyelid and superficial gland formation starts on the 35th day of gestation. On the 50th and 63rd day of gestation, the gland surrounding the cartilage of the third eyelid is composed of the high amount of loose connective tissue and gland cells which give rise to excretory segments. On the 94th day of gestation, the gland lobes become visible, the efferent ducts form. On the 112th day, the cartilage of the third eyelid assumes the appearance of the mature hyaline cartilage. The excretory segments are composed of simple cuboid epithelium with a large, round nucleus arranged less or more peripherally. Their number increases 2- or even 3-fold at the end of gestation.     

Mycobacterial infection of pigs in Croatia

During a five-year period (2000 to 2004) 74,342 pigs were tested by the intradermal tuberculin test in Croatia. Of them, 248 (0.33%) pigs were positive and 91 (0.12%) were found to be suspicious in 7 out of the 13 farms included in the study. Gross pathological changes characteristic of tuberculosis were observed in tuberculin-positive and/or suspicious swine. Mycobacterium was isolated from the lymph nodes of 183 out of 234 swine (78.2%). For better epidemiological understanding, isolates were typed by conventional methods, PCR and hybridisation. The results show that most of the isolates belonged to the Mycobacterium avium complex (175 isolates, 95.7%). Other isolates belonged to M. fortuitum (6 isolates, 3.3%), M. chelonae (1 isolate, 0.5%), and M. peregrinum (1 isolate, 0.5%). Isolated strains of the M. avium complex were identified as M a. avium (37 isolates, 21.1%) and M. a. hominissuis (138 isolates, 78.9%).     

First description of postweaning multisystemic wasting syndrome (PMWS) in wild boar (Sus scrofa) in Croatia and phylogenetic analysis of partial PCV2 sequences

This report describes the first case of postweaning multisystemic wasting syndrome (PMWS) in wild boar in Croatia. During the winter season of 2004, eight wild young piglets (of approximately 2 to 5 months of age) were found dead in a fenced hunting area. Polymerase chain reaction (PCR) was carried out on mesenteric lymph nodes and all animals yielded positive results. In one of these animals diagnosis of PMWS was established based on the three key diagnostic criteria including the clinical manifestation, moderate lymphoid lesions consisting of lymphocyte depletion and granulomatous inflammation, and detection of the presence of PCV2 genome within the lymphoid lesions by in situ hybridisation (ISH). Three additional wild piglets had also mild PMWS-like lesions and a low amount of PCV2 was also found. No PMWS-like lesions or PCV2 genome were detected in the rest of the wild piglets studied. Three PCR-positive isolates were partially sequenced, which confirmed the diagnosis of PCV2 and demonstrated that the three sequences were genetically identical. The phylogenetic analysis of a representative PCV2 isolate indicated that its sequence (DQ875444) is grouped in a separate branch with Hungarian isolate (AY256460) and differs from any of the annotated sequences.     

A serological study on Brucella abortus, caprine arthritis-encephalitis virus and Leptospira in dairy goats in Rio de Janeiro, Brazil

In spite of the large number of goats found in several developing tropical countries, milk production remains unsatisfactory. The occurrence of infectious diseases, such as leptospirosis, brucellosis and caprine arthritis-encephalitis (CAE) may in part be responsible for sub-optimal production. In this study, 1000 serum samples were tested for leptospirosis, 953 for brucellosis and 562 for CAE. All tested flocks presented at least one seroreactive animal for leptospirosis and for CAE. Reactivity to leptospirosis was 11.1%, and serovar hardjo was the most frequently found. Anti-B. abortus agglutinins were found in 0.5% of the samples presented and 14.1% were seroreactive to CAE. Leptospirosis was considered to represent the major infectious problem in the studied goat flocks. The occurrence of infectious diseases in the tested flocks may represent an important factor contributing to the decreased productivity of the animals. These findings may be similar to those observed in other developing countries and require further study to define the relationship between seropositivity and reduced production.     

Nutritive quality and energy yield of high oil,opaque-2 and waxy maize hybrids compared to normal maize hybrids

Grain yield, chemical composition, protein quality and digestible energy were measured for 18 maize varieties (opaque-2, high oil, waxy hybrids, normal hybrids) cultivated in Yugoslavia. Protein quality assessment was based on amino acid composition, and true protein digestibility, biological value, net protein utilization and utilizable protein as determined in nitrogen balance experiments with rats. The protein concentration in dry matter varied from 8.63–18.81% while the fat concentration varied from 4.45–16.13%. There were only minor differences in ash and crude fibre, while starch and sugar varied inversely to fat and protein concentration. Lysine levels were approximately 30% higher for the opaque-2 varieties compared with normal maize, while leucine levels were about 30% lower. Yields were extremely high for all varieties with the highest value being 10.3 tonnes/ha. Due to the high yields, protein production/ha was high in range of 822–977 kg. This resulted in high yields of amino acids/ha. Protein utilization was very high in the opaque-2 varieties, which had the highest lysine contents. The biological values were close to 80 in these varieties, whereas they were only 60–65 in the normal maize varieties.     

Antioxidant and Antihypertensive Activity of Extract from Thymus serpyllum L. in Experimental Hypertension

The low incidence of cardiovascular disease in Mediterranean countries leads to an increased interest of the scientific community for the Mediterranean diet. Our aim was to evaluate total phenol and flavonoid contents, antioxidant capacity, free radical scavenging activity and potential antihypertensive effect of aqueous extract obtained from Thymus serpyllum L. (wild thyme, TE), an aromatic herb from the Lamiaceae family (highly present in Mediterranean diet), in spontaneously hypertensive rats (SHR) and in normotensive Wistar rats. Total phenol content of TE was 2008.33?±?10.6 mg/L GAE, and rosmarinic and caffeic acids were predominant phenolic compounds. The ferric reducing/antioxidant power and antioxidant capacity analysis revealed strong antioxidative properties of TE. In vitro nitric oxide-scavenging activity of 1 mg/l TE was 63.43 % with the IC₅₀ value of 122.36 μg/ml. Bolus injection of TE (100 mg/kg body weight i.v.) induced significant decrease of systolic and diastolic blood pressure and total peripheral resistance in SHR, without effects on these parameters in normotensive Wistar rats. Cardiac index remained unchanged after TE treatment in all experimental rats. Given dose of TE did not show significant nitric oxide-scavenging activity in vivo. Our results indicate that TE may protect against hypertension in experimental model of essential hypertension.     

Heavy Metal Content of Soils and Plum Orchards in an Uncontaminated Area

There are multiple stressors derived from urbanizations that result in frequent disturbances on streams and rivers reducing water quality and threatens aquatic biota. P-glycoprotein (P-gp)-mediated multixenobiotic resistance (MXR) is a defence mechanism analogous to multidrug resistance (MDR), which has been demonstrated in several aquatic organisms. This system protects cells against the entry and the accumulation of xenobiotics and has been proposed as a biomarker for pollution assessment. We conducted a study in a post-urban reach of Esquel stream (Chubut Province) downstream a wastewater treatment plant, in order to assess the presence and activity of MXR in five freshwater macroinvertebrate species (Helobdella michaelseni, Helobdella simplex, Patagoniobdella variabilis, Hyalella curvispina and Chironomus riparius). We measured the accumulation of the model P-gp substrate rhodamine B (RB) in organisms previously exposed to pollution. Our results described the activity of the MXR system in the three species of leeches suggesting their suitability as the in vivo biomonitoring. We also identified a dependence of the transporter activity with the development stage in H. simplex, highlighting the importance of using organisms of similar size classes since it may affect observed results. Finally, we concluded that benthic freshwater macroinvertebrates possess different species-specific levels of MXR activity possibly influencing their natural distribution as well as their survival.     

Phage displayed peptides and anti-idiotype antibodies recognised by a monoclonal antibody directed against a diagnostic antigen of Mycoplasma capricolum subsp. capripneumoniae

A monoclonal antibody (Mab 4.52) raised against Mycoplasma capricolum subsp. capripneumoniae (Mccp) cell lysate was used as a template to obtain substitute antigens recognised by its paratope. Two approaches were investigated: a 17-mer random peptide library displayed on the surface of a filamentous phage was screened by panning on the immobilised Mab 4.52 and anti-idiotype antibodies were generated by immunising a chicken with the F(ab')(2) fragments of the antibody. Analysis of the peptide sequences displayed by the isolated phages identified two peptides. Both contained two cysteine residues and had identical or similar amino acids in positions 5 (P), 8 (I/L) and 13 (L). The fusion phages were also recognised by Mab 4.52 in enzyme-linked immunosorbent assay (ELISA) and binding was shown by surface plasmon resonance. One of the peptides was a markedly better inhibitor (67%) of the binding of Mab 4.52 to its original antigen than the other (20%) at 1mg/ml. After absorption, to remove isotypic and allotypic reactivities, the anti-idiotype IgY was specifically recognised by Mab 4.52 in ELISA and was able to inhibit its binding to the original antigen, whereas anti-idiotype antibodies raised against a bluetongue virus-specific antibody had no effect. In spite of unequivocal binding of the anti-idiotype antibodies and the fusion phages to the paratope of Mab 4.52, goat antisera appeared not to react with either of the surrogate antigens. In contrast, the test sera bound to the original antigen suggesting that Mab 4.52 does not recognise exactly the same antigenic site as antibodies in the goat antisera.     

Trichinella pseudospiralis in pig from Croatia

Trichinella spiralis and Trichinella britovi are species that are frequently found in domestic pigs and various sylvatic animals in Croatia. During routine trichinoscopy, non-encapsulated larvae were detected in the muscle tissue of a domestic pig. Artificial digestion revealed a larvae burden of 602 muscle larvae per gram of tissue. Tissue section analysis confirmed the presence of non-encapsulated larvae. Multiplex PCR identified the larvae as T. pseudospiralis. This observation is consistent with the reports of a local veterinary inspector who described the presence of non-encapsulated Trichinella in four individual cases over the last 2 years. This is the first report of T. pseudospiralis in Croatia and one of very few cases of T. pseudospiralis infection described in domestic pigs. The detection of non-encapsulated larvae stresses the need for implementation of artificial digestion instead of trichinoscopy for the detection and identification of Trichinella infections.