Establishment of a leukocyte cell line derived from peritoneal macrophages of fish,Labeo rohita (Hamilton, 1822)

A continuous leukocyte cell line with phagocytic activity was established from peritoneal macrophages of rohu, Labeo rohita (LRPM). LRPM was initiated from adherent mononuclear leukocytes isolated from peritoneal cavity of rohu, without use of any growth factors or feeder cells. These cells exhibited maximum growth at 30 °C in L-15 medium containing 20 % foetal bovine serum, and has been subcultured for more than 60 passages till date. The cells showed 85 % viability after 6 months of storage in liquid nitrogen. The species of origin of the LRPM was confirmed by the amplification and sequencing of 655 bp fragment of cytochrome oxidase subunit I of mitochondrial DNA. Functionally, LRPM showed phagocytic activity of yeast cells and fluorescent latex beads as evaluated by phase contrast and scanning electron microscopy, respectively. Immuno-modulators such as bacterial lipopolysaccharide and phorbol myristate acetate resulted in functional activation of LRPM; and enhanced their microbicidal activity through release of reactive oxygen species and nitric oxide. Culture supernatant from activated cells also revealed lysozyme activity. Cells of LRPM were positive for alpha-naphthyl acetate esterase enzyme indicating macrophage lineage. Our results indicate that this cell line can be a useful in vitro tool to study the role of macrophages in teleost immune system and to evaluate the effects of new aquaculture drugs. The LRPM cell line represents the first reported leukocyte cell line of peritoneal origin from any freshwater species of fish.     

Nutritional evaluation of fermented black gram (Phaseolus mungo) seed meal in compound diets for rohu, Labeo rohita (Hamilton), fingerlings

Six isonitrogenous (approximately 35% crude protein) and isocaloric (approximately 4.0 kcal g⁻¹) diets were formulated incorporating raw and fermented black gram, Phaseolus mungo, seed meal at 20%, 30% and 40% levels by weight into a fishmeal‐based control diet fed to rohu, Labeo rohita, fingerlings (mean weight, 1.81 ± 0.21 g) for 80 days for a study of fish performance. A particular bacterial strain (Bacillus sp.) isolated from the intestine of adult common carp (Cyprinus carpio) reared in the wild having significant amylolytic, cellulolytic, lipolytic and proteolytic activities was used for fermentation of seed meal for 15 days at 37 ± 2°C. Fermentation of P. mungo seed meal was effective in significantly reducing the crude fibre content and antinutritional factors such as tannins and phytic acid, and enhancing available free amino acids and fatty acids. In terms of growth, feed conversion ratio and protein efficiency ratio, the 30% fermented black gram seed meal incorporated diet resulted in a significantly (P < 0.05) better performance of rohu fingerlings. In general, growth and feed utilization efficiencies of diets containing fermented seed meal were superior to diets containing raw seed meal. The apparent protein digestibility (APD) values decreased with increasing levels of raw seed meal in the diets. The APD for raw seed meal was lower at all levels of inclusion in comparison to those for the fermented seed meals. The maximum deposition of protein in the carcass was recorded in fish fed the diet containing 40% fermented seed meal. The results indicate that fermented black gram seed meal can be incorporated in carp diets up to the 30% level compared to the 10% level of raw seed meal.     

Stock structure analysis of Labeo rohita (Hamilton, 1822) across the Ganga basin (India) using a truss network system

In this study the intraspecific variation of wild Labeo rohita was investigated on the basis of morphometric characters using the truss network system constructed from the fish body. Altogether 435 fish samples were collected from six drainages of the Ganga basin in India. Data were subjected to principal component analysis, discriminant function analysis and univariate analysis of variance. The first principal component (PC1) explained 47.88% of the total variation, while PC2 and PC3 explained 17.22 and 8.33%, respectively. The step‐wise discriminant function analysis retained three variables that significantly discriminated the populations. Using these variables, 62.3% of the original groups were classified into their correct samples and 53.1% of the cross‐validated groups omitting one procedure were classified into their correct samples. Misclassification was higher for samples from the River Gomti (28.6%). Of the total of 31 transformed truss measurements, 30 exhibited significant differences among populations. These findings indicate the presence of six different stocks of L. rohita in the Ganga basin.     

Development and characterization of a continuous macrophage cell line,LRTM, derived from thymus of Labeo rohita (Hamilton 1822)

A long-term thymic macrophage cell line from the thymus explants of Labeo rohita designated as LRTM (L. rohita thymic macrophages) was established, which has been maintained in culture for more than 1 yr. This cell line designated LRTM cells have been subcultured for 70 passages. The cells shape was initially long and elongated; with subsequent passages, the cells became short and epithelial like. The cells exhibited optimum growth in L-15 containing 10% fetal bovine serum and also in Dulbecco’s modified Eagle’s medium at 37°C with 5% CO₂ and showed 85+?% viability after 12 mo storage in liquid nitrogen. In addition, cells showed nonspecific esterase and surface expression of Fc receptors for immunoglobulin G and classes I and II major histocompatibility complex antigens. These observations confirmed that this cell line had the morphologic and functional features as a macrophage. The cells exhibited phagocytic activity by engulfing yeast cells as well as fluorescent latex beads, which was demonstrated by scanning electron microscopy and Giemsa staining. The long-term cultured cells show rapid production of reactive oxygen and nitrogen intermediates following stimulation with lipopolysaccharides and phorbol miristate acetate (PMA). Mostly, all the cells were alpha napthyl esterase acetate positive. After stimulation with PMA and lipopolysaccharide, cultured fish macrophages produced reactive oxygen and nitrogen intermediates. The karyotype analysis showed that these cells have a tetraploid karyotype with 100 chromosomes in each cell, indicating that they are normal L. rohita cells. Amplification, sequencing, and alignment of fragments of two mitochondrial genes 12S rRNA from rohu confirmed that the cell line originated from L. rohita. This cell line should be useful for studying the role of thymic macrophages in differentiation and maturation of thymocytes and can be source of macrophage-specific enzymes and cytokines. The macrophage cell line will be invaluable in studies of pathogen/macrophage interactions, the mechanisms of macrophage antimicrobial effector functions and the contribution of macrophages to the specific immune responses of teleosts.     

Myxobolus insignis sp. n. (Myxozoa, Myxosporea, Myxobolidae), a parasite of the Amazonian teleost fish Semaprochilodus insignis (Osteichthyes, Prochilodontidae)

A new myxosporean species is described from the fish Semaprochilodus insignis captured from the Amazon River, near Manaus. Myxobolus insignis sp. n. was located in the gills of the host forming plasmodia inside the secondary gill lamellae. The spores had a thick wall (1.5-2 microm) all around their body, and the valves were symmetrical and smooth. The spores were a little longer than wide, with rounded extremities, in frontal view, and oval in lateral view. They were 14.5 (14-15) microm long by 11.3 (11-12) microm wide and 7.8 (7-8) microm thick. Some spores showed the presence of a triangular thickening of the internal face of the wall near the posterior end of the polar capsules. This thickening could occur in one of the sides of the spore or in both sides. The polar capsules were large and equal in size surpassing the midlength of the spore. They were oval with the posterior extremity rounded, and converging anteriorly with tapered ends. They were 7.6 (7-8) microm long by 4.2 (3-5) microm wide, and the polar filament formed 6 coils slightly obliquely to the axis of the polar capsule. An intercapsular appendix was present. There was no mucous envelope or distinct iodinophilous vacuole.     

Myxobolus desaequalis n. sp. (Myxozoa,Myxosporea), parasite of the Amazonian freshwater fish,Apteronotus albifrons (Teleostei,Apteronotidae)

Myxobolus desaequalis n. sp. is described from the gill lamellae of the freshwater fish Apteronotus albifrons, collected in the Amazon River, near the city of Salvaterra, Brazil. Large spherical plasmodia filled with disporic pansporoblasts and spores were observed. Ellipsoidal to pyriform spores are 18.3 microm length x 11.2 microm width x 4.4 microm thickness. The anterior end of the spores contain two extremely unequal pyriform polar capsules measuring: (larger): 11.2 microm length, 4.9 microm width, and an isofilar polar filament with 11 to 12 turns obliquely to the longitudinal axis; (smaller): 4.6 microm length, 2.8 microm width, and an isofilar polar filament with 4 to 5 turns, obliquely to the longitudinal axis.     

Myxobolus absonus sp. n. (Myxozoa: Myxosporea) parasitizing pimelodus maculatus (siluriformes: Pimelodidae), a South American freshwater fish

A new myxoporean species is described from a freshwater fish in Brazil. Myxobolus absonus sp. n. was found infecting Pimelodus maculatus captured in the river Piracicaba, State of S?o Paulo, Brazil. Cysts were found free in the opercular cavity. The spores are large (length--15.7 +/- 1.5 microns, width--10.2 +/- 0.7 microns; mean +/- S.D.) and oval in shape, with the anterior end slightly pointed. The spore valves are relatively thin, smooth, and asymmetrical in a frontal view. The polar capsules are pyriform in shape, and unequal in size; the largest are 6.4 +/- 0.7 microns long and 3.6 +/- 0.5 microns wide, while the smallest are 4.2 +/- 0.6 microns long and 2.5 +/- 0.5 microns wide.     

Myxobolus goensis n. sp. (Myxozoa, Myxosporea, Myxobolidae), a parasite of the gills of Mugil cephalus (Osteichthyes, Mugilidae) from Goa, India

Two species of Myxobolus are reported from the gills of Mugil cephalus collected at Goa, India: M. goensis n. sp. and M. parvus Shulman, 1962. Myxobolus goensis n. sp. forms digitiform or rounded plasmodia between the gill rakers. Their spores are oval in frontal view, with tapered anterior extremity, and lemon-shaped in lateral view, measuring 9.7 (9.5-10.5) microm in length, 6.6 (6-7.5) microm in width, and 5.2 (5-6) microm in thickness. The polar capsules are pyriform and unequal in size. The larger ones are 5.3 (4.5-6) microm long and 2.4 (2-3) microm wide; the smaller ones are 2.4 (2-3) microm long and 1.8 (1.5-2) microm wide. The polar filament forms five turns aligned perpendicularly to the longitudinal axis of the larger polar capsules. Within the smaller polar capsules the polar filament is difficult to observe and, apparently, forms three coils. The spores are distinctly different from other Myxobolus species infecting M. cephalus and other Mugil spp. Furthermore, the present material is also different from 204 Myxobolus species presenting differently sized polar capsules, representing nearly all the known species with this characteristic. The fact that only the M. cephalus specimens were infected among a sample of 206 fish specimens, comprising 27 different species, strongly suggests that this parasite is specific to M. cephalus.     

Improvement of nutritive value of grass pea (Lathyrus sativus) seed meal in the formulated diets for rohu, Labeo rohita (Hamilton) fingerlings after fermentation with a fish gut bacterium

Eight isonitrogenous (35% crude protein approximately) and isocaloric (4.0 kcalg(-1) approximately) diets were formulated incorporating raw and fermented grass pea (Lathyrus sativus) seed meal at 10%, 20%, 30% and 40% levels by weight into a fish meal based diet and fed to rohu, Labeo rohita, fingerlings for 80 days and fish performance was studied. A particular bacterial strain (Bacillus sp.) isolated from the intestine of adult common carp (Cyprinus carpio) reared in the wild having significant amylolytic, cellulolytic, lipolytic and proteolytic activities were used for fermentation of seed meal for 15 days at 37 degrees C. Fermentation of grass pea seed meal was effective in significantly reducing the crude fibre content and anti-nutritional factors, such as tannins, phytic acid and the neurotoxin, beta-ODAP and enhancing the available free amino acids and fatty acids. In terms of growth response, feed conversion ratio and protein efficiency ratio, 30% fermented grass pea seed meal incorporated diet resulted in significantly (P < 0.05) better performance of rohu fingerlings. In general, growth and feed utilization efficiencies of fish fed diets containing fermented seed meal were superior to those fed diets containing raw seed meal. The apparent protein digestibility (APD) values decreased with increasing levels of raw seed meal in the diets. The APD for raw seed meal was lower at all levels of inclusion in comparison to those for the fermented seed meals. The highest deposition of carcass protein was recorded in fish fed the diet containing 40% fermented seed meal. The results indicated that fermented grass pea seed meal can be incorporated in carp diets up to 30% level compared to 10% level of raw seed meal.     

Morphological,ultrastructural and phylogenetic analyses of Myxobolus hilarii n. sp. (Myxozoa,Myxosporea), a renal parasite of farmed Brycon hilarii in Brazil

Myxobolus hilarii n. sp. was described, based on morphology, histology, ultrastructure and 18S rDNA sequencing, infecting the kidney of Brycon hilarii (Valenciennes 1850) (Characiformes: Bryconidae) taken from fish farms in the state of São Paulo, Brazil. Thirteen specimens of B. hilarii were examined and 100% had round, white plasmodia in the kidney. The mature myxospores were rounded, measuring 11.5 ± 0.8 (9.8–13.4) μm in length, 11.0 ± 0.7 (9.7–12.4) μm in width and 7.6 ± 1.0 (6.7–9.0) μm in thickness. Polar capsules were elongated and of equal size, with 6.5 ± 0.4 (6.0–7.2) μm in length and 4.0 ± 0.2 (3.6–5.3) μm in width and their polar filaments had 5 to 7 coils. Histological analysis revealed plasmodial development in the renal tubules, causing compression and deformation of adjacent tissues and destruction of renal tubule cells. Ultrastructural analysis showed direct contact between the plasmodial wall and the host tissue and asynchronous plasmodial development. The phylogenetic analysis of South American myxobolids, based on 18S rDNA sequencing, showed the myxosporeans grouping into two main clades. M. hilarii n. sp. appears as sister species of Myxobolus piraputangae.     

条鳅科中国新记录种：曼德勒副棘鳅

副棘鳅属(Paracanthocobitis)隶属于鲤形目(Cypriniformes)条鳅科(Nemacheilidae),分布范围西起巴基斯坦的印度河流域,东至老挝和柬埔寨的湄公河流域。2022年7月,在我国云南省德宏傣族景颇族自治州盈江县采集到一批副棘鳅属鱼类标本,经形态性状和分子系统比较,确认为曼德勒副棘鳅(P. mandalayensis),为中国新记录种。本文对中国境内曼德勒副棘鳅进行了形态描述并提供了物种鉴定的分子依据：该种的主要特征为下唇中央部分有一对中断而膨大的片状结构,密布乳突;尾柄基部上缘有一眼状斑;雄性具有眶下沟;侧线完全;腹鳍基部具腋鳞;背鳍有10?分枝鳍条;尾鳍有4 ~ 8条黑色条带。此外,基于系统发育树和间断地理分布推测,曼德勒副棘鳅种内可能存在隐存种,同时对沙副棘鳅(P. botia)中国分布的记录提出了质疑。这一研究为高黎贡山地区的鱼类资源调查和保护提供了科学依据,并对东南亚地区副棘鳅属物种的鉴定提供了新的依据。     

Henneguya mbakaouensis sp. nov., Myxobolus nounensis sp. nov. and M. hydrocyni Kostoingue & Toguebaye, 1994, Myxosporea (Myxozoa) parasites of Centropomidae, Cichlidae and Characidae (Teleosts) of the Sanaga basin in Cameroon (Central Africa)

The study of 102 teleost freshwater fishes of Sanaga basin in Cameroon revealed the presence of three myxosporean species, among which two were new. Host fishes were of three families: Centropomidae, Cichlidae and Characidae. New species were identified as Henneguya mbakaouensis sp. nov., a gill parasite of Lates niloticus and Myxobolus nounensis sp. nov. found in the kidney and spleen of Sarotherodon galilaeus and Tilapia mariae. Myxobolus hydrocyni Kosto?ngue & Toguebaye, 1994, previously described in Chad, was also found in Cameroon; complementary informations were given on that parasite which seemed to be specific to its host.     

一种硅藻中国新记录属种——科氏杜氏藻

科氏杜氏藻是一种泥生或附沙生硅藻。该文对最近刚确立的一个硅藻属——杜氏藻属进行了介绍,利用光学和扫描电子显微镜对2019年4月20日采自东洞庭湖国家级自然保护区的科氏杜氏藻标本进行了观察研究。结果表明该标本具有以下主要形态特征:(1)壳面椭圆披针形。(2)中央区为蝴蝶结形,没有延伸至壳缘。(3)线纹在壳面大部分区域呈辐射状排列,在两端近平行排列,中部线纹密度20~22条/10μm。(4)孔纹圆形或近圆形,在内壳面被圆顶状的孔膜覆盖。(5)在壳面两端存在假隔片。该文调查采集的科氏杜氏藻种群的形态特征与模式种群相吻合,该属在中国是首次报道,为中国新记录属。     

Sulfomucins and sialomucins in the mucus cells of the alimentary canal in an Indian freshwater major carp, Labeo rohita (Hamilton): a fluorescence microscopic study

The localization of sulfomucins and sialomucins present in the various types of mucus cells--secretory (SMC) and mature non-secretory mucus cells (MMC) of the different regions of the alimentary canal in Labeo rohita (Ham.) has been studied by fluorescence microscopical techniques (viz., Ferric alum Coriphosphine--FAC, Coriphosphine thiazol yellow--CTY and Dische reaction--DR). High degree of fluorescence activity confirms the presence of relatively high amount of sulfomucins in the mucus cells of the oesophageal and buccopharyngeal regions when tested with FAC and CTY methods while moderate intensity of the same in the mucus cells of the two aforesaid regions indicates the localization of relatively lesser amount of sialomucins studied by DR technique. Moderate degree of fluorescence is also observed in the mucus cells (in both SMC and MMC) of the intestinal region with the techniques of FAC, CTY and DR whereas weak fluorescence of FAC, CTY and DR methods is marked in the mucus cells (in both SMC and MMC) of the rectal region due to the presence of low amount of sulfomucins and sialomucins.     

中国翼手类新记录--小巨足蝠

2003年9月在云南获得1号雌性蝙蝠标本,经鉴定为中国新记录——小巨足蝠(Myotis hasseltii),标本保存在中国科学院动物研究所。     

中国翼手目新记录——马来假吸血蝠

2006年11月26日在云南省勐腊县勐仑镇翠鼻峰一无名洞捕捉到1只雄性蝙蝠,体型中等,前臂长60.7 mm,体重21.4 g。耳大,椭圆形,两耳于前额上方约15%的高度相连,耳屏细长双叉;鼻叶较简单,后鼻叶椭圆形,顶部钝圆、两侧缘向外隆凸,中央具一条显著的纵形隆脊,隆脊下部与隆起的间鼻叶相接,间鼻叶呈三角形,顶部W-形,前鼻叶呈马蹄状,前端紧接吻部;无尾,第二指仅具第一指骨;上门齿缺失,上犬齿前后基部均有一较大的副尖,Pm³缺失,下颚第二前臼齿（Pm₃）缺失。经鉴定,该蝙蝠为中国新记录——马来假吸血蝠（Megaderma spasma）。该文给出此蝙蝠的形态与头骨测量数据,并与国内分布的另一种假吸血蝠——印度假吸血蝠（M. lyra）进行了对比。标本保存在广东省昆虫研究所。     

Responses of Corpuscles of Stannius to intra-peritoneal vitamin-D3 administration in teleost Labeo rohita (Hamilton, 1822) reared in water with two different levels of calcium concentration

This study assessed the responses of vitamin-D₃ intraperitoneally injected to Rohu, Labeo rohita @ of 0 IU/kg bw (only solvent), 100 IU/kg bw and 500 IU/kg bw reared in 20 and 40 ppm of calcium (Ca) enriched water. The cellular changes in Corpuscles of Stannius (CS) gland, serum Ca, and inorganic phosphate (P_i) level were analysed up to the 60th day. Rohu administered with 100 IU/kg bw D₃ and exposed to 40 ppm Ca-rich water exhibited notable hyperplasia of CS compared with their control groups. Notable changes with high serum Ca level (13.87 ± 0.3 mg/dl) was detected on the 5th day in fish exposed to 40 ppm Ca-rich water, while related values attained (13.74 ± 0.1 mg/dl) only after 7 days in 20 ppm Ca-rich water of 500 IU/kg bw vitamin D₃ injection. Similarly, high serum P_i level (7.66 ± 0.2 mg/dl) in 40 ppm Ca injected with D₃ at 500 IU/kg bw. The results demonstrated that the Ca homeostasis of Labeo rohita is influenced by intra-peritoneal vitamin D₃. Progressive studies should be conducted by increasing the dose of vitamin D₃ to investigate optimum dose/supplement in feed for commercially important aquaculture teleost Labeo rohita for maximum and sustainable absorption of Ca from the variable water Calcium levels to maintain Ca²⁺ homeostasis.     

Choanoflagellates (Sarcomastigophora, Protozoa) from the Coastal Waters of Taiwan and Japan. I. Three New Species

Three new acanthoecidaean species collected from the coastal waters of Taiwan and Japan are described: Acanthocorbis camarensis n. sp. resembles Acanthocorbis unguiculata in lorica morphology, but differs in having regularly arranged longitudinal and transverse costae at the anterior lorica chamber, and in lacking a nail at the apical end of anterior spine; Diaphanoeca spiralifurca n. sp. is characterized by the spiral arrangement of the costal strips in the posterior half of the lorica chamber and is closely related to Diaphanoeca grandis; Stephanoeca supracostata n. sp. is closely related to Stephanoeca elegans, but differs in having an additional transverse costa at the anterior lorica chamber.     

中国盗猎蝽属一新记录种——红缘盗猎蝽(半翅目:猎蝽科:盗猎蝽亚科)

狭义的盗猎蝽属是猎蝽科盗猎蝽亚科中1个较大的属,已知约40个有效种,主要分布于古北区、东洋区和非洲区,中国已知6种.我们在对新疆半翅目多样性的研究过程中,发现了广布于古北区的红缘盗猎蝽Peirates hybridus(Scopoli,1763).本文重新描述了这一中国新记录种;该种体中型,体表大部分区域黑色,革片、侧...