Moderate pathogenic effect of Ligophorus uruguayense (Monogenoidea, Ancyrocephalidae) in juvenile mullet Mugil liza (Actinopterygii, Mugilidae) from Brazil

ABSTRACT Monogenoidea pathogenic activity can elicit various histological responses in fish. Species of Ligophorus are specific parasites of mullets, and its relationship with host fish may result in a moderate pathogenic action. In order to ascertain this relationship, estuarine mullets (Mugil liza) were collected in an estuary, reared in laboratory, for three weeks, and forwarded for histological and parasitological analyses. Ligophorus uruguayense (Monogenoidea) infestation in the gills of the mullets was identified. The severe infestation by only one species of Monogenoidea may result from the specificity of these parasites to mullets. Mullets submitted to histological analysis exhibited respiratory epithelium detachment; mild, moderate and severe hyperplasia of the respiratory epithelium; atrophy; and telangiectasia of the gills. This is the first study reporting that mullets highly infested by Monogenoidea can show mild (100%) to severe (20%) gill changes with a distinct frequency of occurrence. Because of the high prevalence of mild alterations observed, it is possible to accept that L. uruguayense is moderately pathogenic to M. liza, even during high prevalence and intensity of infestation, as a result of its specificity.

Larval Gnathostoma spinigerum Detected in Asian Swamp Eels, Monopterus albus, Purchased from a Local Market in Yangon, Myanmar

The present study was performed to determine the infection status of swamp eels with Gnathostoma sp. larvae in Myanmar. We purchased total 37 Asian swamp eels, Monopterus albus, from a local market in Yangon in June and December 2013 and 2014. All collected eels were transferred with ice to our laboratory and each of them was examined by the artificial digestion technique. A total of 401 larval gnathostomes (1-96 larvae/eel) were detected in 33 (89.2%) swamp eels. Most of the larvae (n=383; 95.5%) were found in the muscle. The remaining 18 larvae were detected in the viscera. The advanced third-stage larvae (AdL3) were 2.3-4.4 mm long and 0.25-0.425 mm wide. The characteristic head bulb (0.093 x 0.221 mm in average size) with 4 rows of hooklets, muscular long esophagus (1.025 mm), and 2 pairs of cervical sacs (0.574 mm) were observed by light microscopy. The average number of hooklets in the 1st, 2nd, 3rd, and 4th rows was 41, 45, 48, and 51, respectively. As scanning electron microscopic findings, the characteristic 4-5 rows of hooklets on the head bulb, a cervical papilla, tegumental spines regularly arranged in the transverse striations, and an anus were well observed. Based on these morphological characters, they were identified as the AdL3 of Gnathostoma spinigerum. By the present study, it has been confirmed for the first time that Asian swamp eels, M. albus, from Yangon, Myanmar are heavily infected with G. spinigerum larvae.

Positivity and Intensity of Gnathostoma spinigerum Infective Larvae in Farmed and Wild-Caught Swamp Eels in Thailand

From July 2008 to June 2009, livers of the swamp eels (Monopterus alba) were investigated for advanced third-stage larvae (AL3) of Gnathostoma spinigerum. Results revealed that 10.2% (106/1,037) and 20.4% (78/383) of farmed eels from Aranyaprathet District, Sa Kaeo Province and those of wild-caught eels obtained from a market in Min Buri District of Bangkok, Thailand were infected, respectively. The prevalence was high during the rainy and winter seasons. The infection rate abruptly decreased in the beginning of summer. The highest infection rate (13.7%) was observed in September and absence of infection (0%) in March-April in the farmed eels. Whereas, in the wild-caught eels, the highest rate (30.7%) was observed in November, and the rate decreased to the lowest at 6.3% in March. The average no. (mean+/-SE) of AL3 per investigated liver in farmed eels (1.1+/-0.2) was significantly lower (P=0.040) than those in the caught eels (0.2+/-0.03). In addition, the intensity of AL3 recovered from each infected liver varied from 1 to 18 (2.3+/-0.3) in the farmed eels and from 1 to 47 (6.3+/-1.2) in the caught eels, respectively. The AL3 intensity showed significant difference (P=0.011) between these 2 different sources of eels. This is the first observation that farmed eels showed positive findings of G. spinigerum infective larvae. This may affect the standard farming of the culture farm and also present a risk of consuming undercooked eels from the wild-caught and farmed eels.

Prevalence of Zoonotic Metacercariae in Two Species of Grouper, Epinephelus coioides and Epinephelus bleekeri, and Flathead Mullet, Mugil cephalus, in Vietnam

Fishborne zoonotic metacercariae have not been reported from brackish water and marine fish from Vietnam waters although these parasites are common in the country's freshwater fish. Both wild-caught and cultured grouper (Epinephelus coioides and Epinephelus bleekeri), and mullet (Mugil cephalus) from brackish and marine waters locate in Khanh Hoa province in central coastal Vietnam were examined, and found positive for zoonotic trematode metacercariae. From grouper, Heterophyopsis continua and Procerovum varium were recovered. The prevalence of H. continua ranged from 2.0 to 6.0% and that for P. varium ranged from 11.6 to 15.8%. Mullet were infected with Pygidiopsis summa and H. continua both of these species are new records for Vietnam. The prevalence of P. summa in mullet was generally high, ranging from 17.6 to 75.5%, and was significantly higher than the prevalence of H. continua (2.5 to 32.4%). There were no significant differences in the prevalence of metacercariae between grouper from natural or cultured habitats, indicating that the highest risk of infection occurs in the wild-caught state prior to their placement in culture. Further, there was no difference in metacercarial prevalence between the 2 species of grouper. Infected wild-caught seed were only observed from January to October. Monthly variation in prevalence suggests seasonal variation in mullet infections occurs in this region with the highest transmission taking place from October to December. Basic investigations on the ecology and epidemiology of these intestinal flukes need to be carried out to determine their significance as a public health problem and the aspects of their biology that may be vulnerable to control interventions.

A survey of Heterophyes nocens and Pygidiopsis summa metacercariae in mullets and gobies along the coastal areas of the Republic of Korea

The infection status of mullets Mugil cephalus (n = 139) and gobies (n = 35) Acanthogobius flavimanus with metacercariae of Heterophyes nocens and Pygidiopsis summa was examined in 11 western, southern, and eastern coastal areas of the Republic of Korea, using a digestion technique. Heterophyid metacercariae were highly prevalent in mullets from western and southern coastal areas; Shinan-gun (100% for H. nocens and 100% for P. summa), Muan-gun (93% and 100%), Buan-gun (42% and 75%), Seocheon-gun (73% and 53%), Ganghwa-gun (47% and 100%), Sacheon-shi (47% and 77%), and Gangjin-gun (50% and 70%, respectively). Only 1 (10%) of 10 mullets from an eastern coastal area, i.e., Donghae-shi, was positive for P. summa metacercariae. Metacercarial densities were the highest in the trunk of mullets for H. nocens and the gill for P. summa. Gobies from Muan-gun were positive for H. nocens (40%) and P. summa metacercariae (40%), and gobies from Seocheon-gun revealed H. nocens metacercariae (20%). The metacercarial density was remarkably higher in mullets than in gobies. The results revealed that H. nocens and P. summa metacercariae are prevalent in mullets and gobies from coastal areas of the Republic of Korea, and the prevalence and intensity of infection vary according to geographical locality.

A new endemic focus of Heterophyes nocens and other heterophyid infections in a coastal area of Gangjin-gun, Jeollanam-do

To know the prevalence of heterophyid trematodes among inhabitants of a southern coastal village, i.e., Sacho-ri, Gangjin-gun, Jeollanam-do (Province), 82 stool samples were examined on helminth eggs and protozoan cysts using Kato-Katz and formalin-ether sedimentation techniques. Total 33 people (40.2%) were positive for trematodes (Heterophyes nocens; 15 people, Pygidiopsis summa; 3, Metagonimus sp.; 4, Clonorchis sinensis; 7, Gymnophalloides seoi; 6) and/or protozoa (Entamoeba coli; 3). Among intestinal trematode egg positive cases, 17 were treated with praziquantel and their whole diarrheic stools were collected after purgation. Adult flukes of H. nocens (number of specimens=1,294), P. summa (386), Stellantchasmus falcatus (5), Stictodora lari (4), and Heterophyopsis continua (1) were collected using a stereomicroscope. To know the source of human H. nocens infections in this village, metacercarial infections in mullets (10) were examined and most H. nocens metacercariae (101/105, 96.2%) were found in the trunk portion. From above results, the surveyed coastal village has been newly known as an endemic focus of human H. nocens infection and consuming raw mullets was the presumable source of human heterophyid infections.

Gnathostoma infection in fish caught for local consumption in Nakhon Nayok Province, Thailand. II. Deasonal variation in swamp eels.

From August 2000 to August 2001, 1844 swamp eels (Monopterus albus) were purchased from several local markets in Nakhon Nayok Province, Thailand, and examined for the presence of Gnathostoma advanced third-stage larvae. The overall prevalence was 30.1% and the mean number of larvae/eel (infection intensity) was 10.0. The highest infection rate (44.1%) was found in August 2000 and the lowest (10.7%) in March 2001. The greatest mean number of larvae/eel (75.1) was found in August 2000, whereas the fewest (2.3) was in July 2001. It is suggested that the prevalence and intensity of infection decreased within two months after the end of the rainy season and started to rise again about two months after the next rainy season began. A total of 5,532 Gnathostoma larvae were recovered from 555 infected eels, with a maximum number of 698 larvae/eel. The highest rates of Gnathostoma infection according to eel body length and weight were 87.5% in the group 91-100 cm, and 100% in groups of 901-1100 g, respectively. There were significant correlations between eel body lengths and infection rates, body lengths and infection intensities; eel body weights were also significantly correlated with infection rates and infection intensities. It was noted that the longer/ heavier the eels were, the higher would be the infection rates and the greater the infection intensities. Tissue distributions of Gnathostoma larvae in the livers and muscles of swamp eels were as follows: 43.0% of the total number of larvae were found in the muscles and 57.0% were in the liver; 29.7, 51.7, and 18.6% were in the anterior, middle, and posterior parts, respectively; 35.1% were in the dorsal part, while 64.9% were in the ventral part; 9.0, 18.7, 7.4, 20.6, 33.1, and 11.2% were in the anterodorsal, mediodorsal, posterodorsal, anteroventral, medioventral and posteroventral parts, respectively. Of the 5,532 Gnathostoma larvae examined, 1101 (19.9%) were found to possess morphological variants or abnormal cephalic hooklets. The most common unusual feature was that there were few to numerous extra rudimentary hooklets below row 4 and between the 4 rows of hooklets (7.6%), the presence of a fifth row of hooklets (3.5%), abnormal hooklets in any of the 4 rows of hooklets (5.2%), spiral arrangement of the 4 rows of hooklets (1.8%), and larvae having only 3 rows of hooklets (0.3%).

Gnathostoma infection in Nakhon Nayok and Prachin Buri, Central Thailand.

Gnathostoma infection in Nakhon Nayok and Prachin Buri Provinces, Central Thailand, was investigated. The prevalence and intensity of infection of swamp eels were determined; dog fecal samples and fresh-water copepods were examined for evidence of infection. The overall prevalence of eel infection was 38.1% (117/307) in Nakhon Nayok and 24.0% (74/308) in Prachin Buri--the former rate being significantly higher than the latter. Most of the positive Nalkhon Nayok eels (53.8%) harbored only 1-9 larvae; only one eel bore more than 50 larvae. In Prachin Buri, 67.6% of the positive eels harbored 1-9 larvae; again, only one eel bore more than 50 larvae. The mean number of 11.0 +/- 10.4 larvae/eel in Nakhon Nayok was not significantly different from that of Prachin Buri (9.3 +/- 11.4). A total of 1,292 gnathostome larvae were recovered from 307 eels in Nakhon Nayok. Of these, 52.3% had accumulated in the liver and 47.7% had spread throughout the muscles. In eels from Prachin Buri, 50.6% and 49.4% of the total of 688 larvae (from 308 eels) were found in the liver and muscles, respectively. The larvae preferred encysting in ventral of muscles rather than dorsal part; they preferred the middle portion to the anterior and posterior portions. The average length of gnathostome larvae recovered from Nakhon Nayok eels was 4.0 +/- 0.5 mm (range 2.5-5.1 mm) and the average body width was 0.40 +/- 0.05 mm (range 0.29-0.51 mm). Those from eels in Prachin Buri were 3.9 +/- 0.5 mm (range 2.2-5.1 mm) and 0.34 +/- 0.05 mm (range 0.20-0.48 mm), respectively. The mean body length and width of the larvae from eels in Nakhon Nayok were significantly greater than those of the larvae from eels in Prachin Buri. In Ban Phrao, Nakhon Nayok, none of the first 44 fecal specimens examined was positive. Of the second (68) and the third (70) specimens, one (1.5%) and two (2.9%) samples were positive. However, six months after the third fecal collection, no eggs were found. In Tha Ngam, Prachin Buri, no eggs were found in all three batches (109, 115, and 100 fecal samples). A cyclops survey of 4,000-5,000 crustacea from each of two areas (Ban Phrao and Tha Ngam) found no evidence of natural cyclops infection.