Bacterial and pathological study of caudal fin rot in brood stocks of Salmo trutta caspius from the propagation and breeding center of Shahid Bahonar of Kelardasht region

In order to study caudal fin rot with emphasis on Aeromonas hydrophila and Pseudomonas fluorescens in Salmo trutta caspius from the salmonids propagation and breeding center of Shahid Bahonar of kelardasht region, One hundred and eighty brood stocks having fin damage symptoms were chosen. Two bacterial samples from each fish were cultured on Aeromonas and Pseudomonas specific media. Biochemical tests, API2OE identification system and antibiogram test using six antibiotic disks were performed for diagnosing isolates bacteria and finding suitable antibiotic. Thirty samples from caudal fin of damaged fishes were fixed in 10% formalin and 51.tm microscopic sections were prepared using standard scatological methods and then stained by Haematoxylin-Eosin staining method to observe the pathological changes and also Maccallum-Goodpasture staining method to observe the bacterial colonies. In second stage of the study, bacterial samples were taken from thirty brood stocks using similar method at the first stage of sampling. For isolation and biochemical diagnosis of Aeromonas and Pseudormonas genus, the samples were analyzed by molecular research included PCR amplification (using 16S rDNA genes of the genus pseudomonas and 16S-23S rDNA intergenic spacer of the genus Aeromonas) and restriction analysis by four restriction enzymes for each genus. The results of biochemical tests showed that isolated bacteria were belonged to Aeromonas caviae and Aeromonas hydrophila (subspecies anaerogenes), Pseudomonas fluorescens, Pseudomonas putida and Pseudomonas alcaligenes while the results of API2OE identification system showed that the isolated bacteria belonged to Aeromonas hydrophila, Pseudomonas fluorescens, Pseudomonas putida and Pseudomonas aeruginosa. Restriction analysis of Aeromonas samples with Hin6l, Csp6I, Taql, and Tasl revealed three samples were different from others while restriction analysis of Pseudomonas samples with Alul, Hinfl, Rsal, and Trull showed at least five species or biovars. The results of antibiogram test showed all Aeromonas samples were sensitive to Trimethoprim, Chloramphenicol and Nitrofurazone, mostly to Nalidixic acid and Chloramphenicol, while most of samples were resistant to Erythromycin and Oxytetracycline. Pseudomonas samples were only sensitive to Nitrofurazone and mostly resistant to Oxytetracycline, Nalidixic acid, Erythromycin, Trimethoprim and Chloramphenicol. The results of light microscope study showed hyperplasia and spongiosis of the malpigian cells of epidermis, increasing of melanin pigments underlying epidermis; sever necrosis in both epidermis and dermis and also sloughing the epidermis in some cases. Occurrence of clefts through the epithelium, neovascularization, hyperemia and mild inflammatory response in dermis and separation of the fin rays also were observed. No bacterial colonies were found in the sections.

The study of histopathological damage in heart and bulbus arteriosus in rainbow trout

This study investigated the pathological changes of heart and bulbus artrius of rainbow trout breeders in several group of ages and density. The aim of study was to consider the process and the intensity of the heart and bulbus arteriosus damages in accordance to gender, age and stocking density of trout in three fish culture center (Zarghezel, Niyak in Haraz Region,Mazandaran and Espiran in Tabriz city environs). In field research, the all records the feed and feeding type, rate of mortality, stocking density of spawners and per spawners fishes, water chemical and physical specification was screened. Stocking density was considered as the most important stressor. 10 fish specimens from 7 weight groups (less than 90g, 90 to 300g, 300 to 500 g, 500 to 1000g, 1 to 3 kg, 3 to 5 kg, over 5 kg), totally 210 specimens were sampled and heart and bulbus arteriosus were taken. Samples were fixed in 10 % formalin and transferred to pathology laboratory of veterinary faculty of Tabriz Azad University. Histopathological slides and H&E staining were prepared from these samples. In total, 47 male and 73 female samples showed cardiovascular injury (29 cases in extensive system, 41 cases in semi intensive system, 50 cases in intensive system). The most important was damages, edema and hyperemia in spongy layer of atrium and ventricle muscles, but degeneration the muscle fibers, moderate edema , minor vascular damage. Hemorrhage as the effect of severs vascular damage, thrombus, sever inflammation, sever degeneration in muscle fiber, necrosis and fibrose were further pathological changed. The results of this study showed that the severity of damage increased by increasing the age (weight) of fishes. This situation was seen in all three culturing system (extensive system, semi intensive system, recirculation system). Histopathological changes is obviously seen in samples over 500g, therefore the damages were found to be important (P<0.05). Pathological effects and its severity in recirculation system was significantly high (P<0.05). Comparison with two other culturing system, histopathological changed in heart and bulbus arterius between male and female was significantly different.

Separation and identification fresh water leeches of waters resources in Kordistan Province

In the primary studying, known leeches have included into free living and parasitic which the parasitic group, besides of direct impacts like: growth detraction, anemia, making wound in the connecting part with the skin, with making plat for secondary bacterial and molding infections in the toll place, is able to cause to transfer blood flagellate and virus to the fish too. Therefore, by accusing information related to fauna leeches of each area a risk factor from the viewpoint of the possibility of being or accusing transferred diseases by these leeches, one can predict and forehand about them. Freshwater leeches of Iran to present accurate recognition (morphological, molecular) have not been, and there are some limited reports from different parts of the country about them. One of the areas that its leeches have not been identified yet is Kurdistan, By having five latrines and big permanent rivers and 32000 springs and a lot of deep and semi-deep wells and this province is a convenient bed for growing aquatics in the country. Therefore, identifying risk factors for development of aquaculture on water resources is one important factor to access achieving development goals. For recognizing leeches of this province, some samples from 10 stands were token. Samples from under stones, sticking to the fish, turtles, plants and solid substances in the water were separated and after recording their physical characteristics, calming with 10% ethylic alcohol with 10% formalin become fixed and after painting with Carmen acetic acid by standard keys for 7 species of Helobdella stagnalis, Placobdella costata, Hemiclepsis marginata, Erpobdella octoculata, Hirudo medicinalis, Dina lineate lineata have been identified and described. Which Helobdella stagnalis has the highest distribution in the province and the minimum one is Hirudo medicinalis. However, that the data obtained in leeches in Kurdistan is a relatively complete collection in this research, recognizing fauna of these areas needs more studying. The Placobdella costata and Hemiclepsis marginata sticking to the fish were separated among identified species which showed that these are parasites for the fish. The sticking area of those leeches to the skin was accompanied with scales cast, damage to mucous membranes beneath the parasite and bleeding Was associated with Histopathology studying effect includes observing break and disconnection in the leech connecting place to the epithelial layer of epidermis in the skin, destroyed nucleus in skin Epithelial cells with observing necrosis in ulcerative place become of the leech and the sub acute inflammated penetration until acute necrosis with opening in Dermis layer is observable. Kidney of this fish have changes such as: proliferation, like proliferative kidney disease with increasing proliferative glomerular cells and increasing in membranous cells in Capillary corpuscle, observing necrotic cells in haematopoietic tissue of kidney along with increasing in infiltration of leukocyte's cells generally mono nucluars such as lymphocytes and less poly morpho nucluars such as neutrophiles that are symptoms of disorders causing anemia become of nourishing and sucking blood by the leech and creating a chronic kidney infection that originally root is in another place like the skin. Also Hemorrhagic anemia causes losing RBC's is because of using the host blood by the leech. (In this situation, one can see immature RBC red cells in Peripheral blood. To identify potential carriers of the leech to the viruses, after finding them in recorded stands and putting them in 75% ethanol for viruses cause IPN, VHS, IHN, they were tested by PCR that the conclusion of these experiments approved IPN virus in Hemiclepsis marginata and Hirudo medicinalis. This kind of leeches can act like a mechanical carrier and causing spreading the agent of this disease. It is worth mentioning that studying the pathogenicity of this virus for aquaculture sources, mentioned before needs more research. During the study of infected fish with leeches that was done after preparing bloody slides and staining them, no case blood parasites was observed. During a research about infecting fish experimentally to known leeches it become clear that 5 days after being in aquarium including leeches, samples of sticking Hirudo medicinalis leech to the golden carp with scales cast were observed. Including leeches to the fish started with molting the scales in the sticking area in the fish and fish become too uneasy and by rubbing themselves to the malls and things inside the aquarium, tried to separate them. Finally, after around 30 hours, leeches penetrate the skin, feeding from blood and tissue liquids and cause mortality the fish and then they become separated from them. If the corpse of these fish stayed in the aquarium, the Helobdella stagnalis and Erpobdella octoculata would start feeding them.

Study of nephrocalcinosis in rainbow trout (Oncorhynchus mykiss), from fry to brood stock

For survey incidence of nephrocalcinosis in rainbow trout fish, during winter 1385, samplings were performed in three fish farm with different water source: river, spring and recirculation system by using well water. In this survey 5 specimens from each 8 groups and in general 120 specimens of this fish were caught by random sampling, and also amounts of O2, CO2, pH and temperature of water were measured. Then blooding and renal tissue sampling performed that renal samples were fixed in 10% buffered formalin and blood samples after separating serum stored in -200C. Renal specimens transferred to pathology laboratory, pathological slides were prepared and stained by hematoxylin & Eosin method. From 120 specimens, 6 cases of fish represent nephrocalcinosis. Pathologic signs include: renal epithelial necrosis, dilated ureters, dense basophilic materials inside the dilated tubules and cast formation in some renal tubules. From 6 cases of nephrocalcinosis, 3 cases (7/5%) were related to recirculation system and 2 (5%) case were related to river water and 1 (2/5%) case was related to spring water. In survey amount of urea, creatinine and Uric acid between different weight groups, distinguished that difference between creatinine middling in different weight groups were significant and also in between healthy and afflicted fish, significant statistical difference were only in creatinine amount between healthy and afflicted fish in each farm. Amounts of O2, pH and temperature of water in three farms were in normal range and only rate of water CO2 in ponds of recirculatory system were very higher (25 mg/lit) than other farms. May be, this reason led to high number of nephrocalcinosis in recirculatory system, than other farms, nemley 7/5% of fish that caught from this farm .This subject is related to the role of CO2 in creating nephrocalcinosis which is descript in references.