Novel Treatment for Lead Exposure in Children with Autism

The pathogenesis of some cardiovascular diseases (CVDs) has been altered with changes in the balance of certain trace and toxic elements. The aim of the present study was to assess the role of zinc (Zn) and cadmium (Cd) in smoker and nonsmoker male CVD patients (n?=?457) of two age groups (31–45) and (46–60). The both elements were determined in biological samples (scalp hair, blood, and urine) of CVD patients and healthy referents for comparison purpose. The concentrations of Zn and Cd were measured by atomic absorption spectrophotometer prior to microwave-assisted acid digestion. It was observed that the mean values of Cd were significantly higher in the biological samples of smokers CVD as compared to nonsmoker CVD patients, while the level of Zn was lower in both smoker and nonsmoker patients. The concentrations of Zn in whole blood and scalp hair samples were lower in CVD patients as compared to referents (p?>?0.001). Results showed significant changes of levels of Cd and Zn in blood and scalp hair samples of CVD patients when compared with healthy referents, while reverse in the case of urine samples. It was observed that low Zn levels were associated with both smoker and nonsmoker CVD patients, while increased cadmium accumulation was observed in smoker patients as compared to nonsmoker patients (p?>?0.025).     

Analysis of Hair Trace Elements in Children with Autism Spectrum Disorders and Communication Disorders

The primary objective of the present study is analysis of hair trace elements content in children with communication disorder (CD) and autism spectrum disorder (ASD). A total of 99 children from control, CD, and ASD groups (n = 33) were examined. All children were additionally divided into two subgroups according to age. Hair levels of trace elements were assessed using inductively coupled plasma mass spectrometry. The difference was considered significant at p < 0.01. The obtained data demonstrate that children with CD are characterized by significantly increased hair lithium (Li) (96 %; p = 0.008), selenium (Se) (66 %; p < 0.001), arsenic (As) (96 %; p = 0.005), beryllium (Be) (150 %; p < 0.001), and cadmium (Cd) (72 %; p = 0.007) content, being higher than the respective control values. In the ASD group, hair copper (Cu), iodine (I), and Be levels tended to be lower than the control values. In turn, the scalp hair content of Se significantly exceeded the control values (33 %; p = 0.004), whereas the level of iron (Fe) and aluminum (Al) tended to increase. After gradation for age, the most prominent differences in children with CD were detected in the elder group (5–8 years), whereas in the case of ASD—in the younger group (3–4 years old). Taking into account the role of hair as excretory mechanism for certain elements including the toxic ones, it can be proposed that children suffering from ASD are characterized by more profound alteration of metal handling and excretion in comparison to CD.     

Interaction Between Selenium and Mercury in Biological Samples of Pakistani Myocardial Infarction Patients at Different Stages as Related to Controls

It has been speculated that trace elements may a play role in the pathogenesis of heart diseases. In the present study, we aimed to assess the levels of selenium (Se) and mercury (Hg) in biological samples (whole blood, urine, and scalp hair) of myocardial infarction (MI) patients of both genders (age range 45–60 years) at the first, second, and third heart attack (n?=?130), hospitalized in a cardiac ward of a civil hospital of Hyderabad City (Pakistan). For comparison, healthy age-matched referent subjects (n?=?61) of both genders were also selected. Se and Hg in biological samples were measured by electrothermal atomic absorption spectrometry and cold vapor atomic absorption spectrometry, prior to microwave acid digestion, respectively. The validity of the methodology was checked by biological certified reference materials. During this study, 78 % of the 32 registered patients of third MI attack (aged >50 years) died. The concentration of Se was decreased in scalp hair and blood samples of MI patients, while Hg was higher in all biological samples as compared to referent subjects. Se concentration was inversely associated with the risk of MI attacks in both genders. These results add to an increasing body of evidence that Se is a protective element for cardiovascular health.     

Disparities of Selected Metal Levels in the Blood and Scalp Hair of Ischemia Heart Disease Patients and Healthy Subjects

Imbalances in the concentrations of trace metals have become an increasingly recognized source of infirmity worldwide particularly in the development of ischemia heart disease (IHD). Present study is intended to analyze the concentrations of Ca, Cd, Co, Cr, Cu, Fe, K, Li, Mg, Mn, Na, Pb, Sr, and Zn in the blood and scalp hair of the patients and counterpart controls by flame atomic absorption spectrometry after wet-acid digestion. On the average, Cd, Co, Cr, Fe, K, Li, Mn, Na, and Pb revealed significantly elevated concentrations in the blood of the patients compared with the controls (p < 0.05), whereas mean levels of Ca, Cd, Fe, K, Li, Pb, and Sr in the scalp hair were significantly higher in the patients than the controls (p < 0.05). Most of the metals exhibited noticeable disparities in their concentrations based on gender, abode, dietary/smoking habits, and occupations of both donor groups. The correlation study and multivariate statistical analyses revealed some significantly divergent associations and apportionment of the metals in both donor groups. Overall, comparative variations of the metal contents in blood/scalp hair of the patients were significantly different than the controls; thus, evaluation of trace metals status may be indicative of pathological disorders, such as IHD.     

Evaluation of Cadmium, Chromium, Nickel, and Zinc in Biological Samples of Psoriasis Patients Living in Pakistani Cement Factory Area

Psoriasis is a noncontiguous common and chronic skin disorder. The aim of the present study was to compare the level of trace elements cadmium (Cd), chromium (Cr), Nickel (Ni), and zinc (Zn) in biological samples (whole blood, urine, and scalp hair) of psoriasis patients of both gender age ranged (25–55 years) at mild, moderate severe, and more severe stage (n?=?418) living in the vicinity of cement factory. For comparison purposes, healthy age-matched referent subjects, residents of industrial and non-industrial area, of both gender were also selected (n?=?241). The concentrations of trace and toxic elements were measured by atomic absorption spectrophotometer prior to microwave-assisted acid digestion. The validity and accuracy of methodology was checked by using certified reference materials (CRMs) and conventional wet acid digestion method on same CRMs and real samples. The results of this study showed that the mean values of Cd, Cr, Ni, and Pb were significantly higher in scalp hair, blood, and urine samples of mild and severe psoriasis patients as compared to referents (p?<?0.001), while the concentration of Zn was lower in the scalp hair and blood, but higher in the urine samples of psoriasis patients. The deficiency of Zn in psoriasis patients may be undoubtedly caused by the toxic element exposures via cement factory.     

Estimation of Copper and Iron Burden in Biological Samples of Various Stages of Hepatitis C and Liver Cirrhosis Patients

There is accumulative evidence that the metabolism of iron (Fe) and copper (Cu) is altered in human due to infections, indicating that both elements have roles in pathogenesis and progress of viral diseases. In the present study, the correlation of Cu and Fe was evaluate in biological samples (serum and scalp hair) of hepatitis C (hepatitis C virus (HCV)) patients of both genders at different stages. For comparative study, the scalp hair and serum samples of healthy individuals of same age group (30–50 years) and socioeconomic status were collected. The biological samples were analyzed for Fe and Cu by atomic absorption spectroscopy after microwave-assisted acid digestion. The validity and accuracy of methodology were checked by certified reference materials of same matrixes. The levels of Cu and Fe in biological samples were enhanced in hepatic disorder patients, including acute (after diagnosis test, anti-HCV sero-positive) hepatic fibrosis and liver cirrhosis as compared to healthy referents. The difference was significant (p?<?0.01) in the case of liver cirrhotic patients. It was observed that the data of Cu and Fe in referents and patients of both genders had normal distributions. The inter-elemental correlation (r) among Cu vs Fe in serum and scalp hair samples of referents and patients were not significant in both genders (p?>?0.1) except in the first stage of HCV (p?<?0.1). It was concluded that the increase of Cu and Fe content in human body seems to contribute to the development of cirrhosis in patients with viral hepatitis C.     

Interaction Between Essential Elements Selenium and Zinc with Cadmium and Mercury in Samples from Hypertensive Patients

The abnormal metabolism of metal ions plays an important role in health and disease conditions; hence, the studies about them have received much interest. The objective of this study was to evaluate the association between trace and toxic elements zinc (Zn), cadmium (Cd), selenium (Se), and mercury (Hg) in biological samples (scalp hair, blood, and urine) of hypertensive patients (n?=?257), residents of Hyderabad, Pakistan. For comparison purpose, the biological samples of age-matched healthy controls were selected as referents. The concentrations of trace and toxic elements were measured by atomic absorption spectrophotometer prior to microwave-assisted acid digestion. The validity and accuracy of the methodology was checked using certified reference materials and by the conventional wet acid digestion method. The recovery of all studied elements was found in the range of 96.4–99.1 % in certified reference materials. The results of this study showed that the mean values of Cd and Hg were significantly higher in scalp hair, blood, and urine samples of hypertensive patients than in referents (P?<?0.001), whilst the concentrations of Zn and Se were lower in the scalp hair and blood, but higher in the urine samples of hypertensive patients. The deficiency of Zn and Se and the high exposure of toxic metals may be synergistic with risk factors associated with hypertension.     

Bioreduction of hexavalent chromium by <Emphasis Type="Italic">Pseudomonas stutzeri</Emphasis> L1 and <Emphasis Type="Italic">Acinetobacter baumannii</Emphasis> L2

Bioreduction of the very toxic hexavalent chromium ion [Cr(VI)] to the non-toxic trivalent chromium ion [Cr(III)] is a key remediation process in chromium-contaminated sites. In this study, we investigated the bioreduction of Cr(VI) by Pseudomonas stutzeri L1 and Acinetobacter baumannii L2. The optimum pH (5–10), temperature (27, 37 and 60 °C) and initial chromium Cr(VI) concentration (100–1000 mg L^?1) for Cr(VI) reduction by strains L1 and L2 were determined using the diphenylcarbazide method. In the presence of L1 and L2, the bioreduction rate of Cr(VI) was 40–97 and 84–99%, respectively. The bioreduction of Cr(VI) by L2 was higher, reaching up to 84%—than that by L1. The results showed that strain L2 was able to survive even if exposed to 1000 mg L^?1 of Cr(VI) and that this tolerance to the effects of Cr(VI) was linked to the activity of soluble enzyme fractions. Overall, A. baumannii L2 would appear to be a potent Cr(VI)-tolerant candidate for the bioremediation of chromium (VI)-contaminated wastewater effluent.     

Isolation and characterization of chromium(VI)-reducing bacteria from tannery effluents and solid wastes

In the present investigation, five novel Cr(VI) reducing bacteria were isolated from tannery effluents and solid wastes and identified as Kosakonia cowanii MKPF2, Klebsiella pneumonia MKPF5, Acinetobacter gerneri MKPF7, Klebsiella variicola MKPF8 and Serratia marcescens MKPF12 by 16S rDNA gene sequence analysis. The maximum tolerance concentration of Cr(VI) as K₂Cr₂O₇ of the bacterial isolates was varying up to 2000 mg/L. Among the investigated bacterial isolates, A. gerneri MKPF7 was best in terms of reduction rate. The optimum temperatures for growth and Cr(VI) reduction by the bacterial isolates were 35 and 40 °C, respectively except A. gerneri MKPF7 which grew and reduced Cr(VI) optimally at 40 °C. The optimum pH for growth and Cr(VI) reduction by K. cowanii MKPF2, A. gerneri MKPF7 and S. marcescens MKPF12 was 7.0 whereas the optimum pH for growth and Cr(VI) reduction by K. pneumoniae MKPF5 and K. variicola MKPF8 were 7.0, 8.0 and 6.0, 7.0, respectively. All the bacterial isolates showed maximum tolerance against Ni²⁺ and Zn²⁺ whereas minimum tolerance was observed against Hg²⁺ and Cd²⁺. The bacteria isolated in the present study thus can be used as eco-friendly biological expedients for the remediation and detoxification of Cr(VI) from the contaminated environments.     

Comparative analysis of prokaryotic diversity in solar salterns in eastern Anatolia (Turkey)

The prokaryotic communities of four salterns (Bingöl, Fadlum, Kemah, and Tuzlagözü) in Turkey were examined and compared using the cultivation and cultivation-independent methods [fluorescence in situ hybridization (FISH) and 454 pyrosequencing]. FISH analysis with universal probes revealed that feeding waters carried 1.6 × 10²–1.7 × 10³ cells mL^?1, while crystallization ponds carried 3.8 × 10⁶–2.0 × 10⁷ cells mL^?1 that were mostly haloarchaea, including square cells (except for Kemah). High-throughput 16S rRNA-based gene sequencing showed that the most frequent archaeal OTUs in Bingöl, Fadlum, Tuzlagözü, and Kemah samples were affiliated with Haloquadratum (76.8 %), Haloarcula (27.8 %), Halorubrum (49.6 %), and Halonotius (59.8 %), respectively. Bacteroidetes was the dominant bacterial phylum in Bingöl and Fadlum, representing 71.5 and 79.5 % of the bacterial OTUs (respectively), while the most abundant bacterial phylum found in the Kemah saltern was Proteobacteria (79.6 %). The majority of the bacterial OTUs recovered from Tuzlagözü belonged to the Cyanobacteria (35.7 %), Bacteroidetes (35.0 %), and Proteobacteria (25.5 %) phyla. Cultivation studies revealed that the archaeal isolates were closely related to the genera Halobacterium, Haloarcula, and Halorubrum. Bacterial isolates were confined to two phyla, Proteobacteria (Alphaproteobacteria and Gammaproteobacteria classes) and Bacteroidetes. Comparative analysis showed that members of the Euryarchaeota, Bacteroidetes, Proteobacteria, and Cyanobacteria phyla were major inhabitants of the solar salterns.     

Additive and synergistic interactions amongst <Emphasis Type="Italic">Orius laevigatus</Emphasis> (Heteroptera: Anthocoridae), entomopathogens and azadirachtin for controlling western flower thrips (Thysanoptera: Thripidae)

This study evaluated the efficacy of the foliage-dwelling predator Orius laevigatus, soil applied entomopathogens and azadirachtin alone and in combinations for controlling western flower thrips (WFT). Evaluated products were Nemastar^® Steinernema carpocapsae (E-nema) and O. laevigatus (Re-natur), Metarhizium anisopliae isolate ICIPE-69 and NeemAzal-T (azadirachtin) (Trifolio). Efficacy against WFT was significantly improved by combined treatments achieving 62–97 % reduction in WFT emergence, compared to 45–74 % in single treatments, and interactions resulted in two synergistic and eight additive responses. Metarhizium-based treatments reduced WFT survival by 93–99.6 % when late mortality by mycosis was considered. Halving the number of released predators did not significantly reduce efficacy (86–96 vs. 76–88 % thrips reduction), and when Orius was introduced to target L1 of WFT, 96–98 % reduction was achieved while only 71–89 % for L2. Early release of O. laevigatus and combination with soil application of NeemAzal-T and/or entomopathogens can be a successful and reliable biocontrol strategy for WFT.     

Concentration of Zinc,Copper, Iron,Calcium, and Magnesium in the Serum,Tissues, and Urine of Streptozotocin-Induced Mild Diabetic Rat Model

A survey carried out in Lake Tana in 2015 found that Hg levels in some fish species exceeded internationally accepted safe levels for fish consumption. The current study assesses human exposure to Hg through fish consumption around the Lake Tana. Of particular interest was that a dietary intake of fishes is currently a health risk for Bihar Dar residents and anglers. Hair samples were collected from three different groups: anglers, college students and teachers, and daily laborers. A questionary includes gender, age, weight, activity. Frequency of fish consumption and origin of the eaten fish were completed by each participant. Mercury concentrations in hair were significantly higher (P value <0.05) for anglers (mean?±?standard deviation 0.120?±?0.199 μg/g) than college students (mean?±?standard deviation 0.018?±?0.039 μg/g) or daily workers (mean?±?standard deviation 16?±?9.5 ng/g). Anglers consumed fish more often than daily workers and college group. Moreover, there was also a strong correlation (P value <0.05) between the logarithms of total mercury and age associated with mercury concentration in scalp hair. Mercury concentrations in the hair of men were on average twice the value of the women. Also, users of skin lightening soap on a daily basis had 2.5 times greater mercury in scalp hair than non-users. Despite the different sources of mercury exposure mentioned above, the mercury concentrations of the scalp hair of participants of this study were below levels deemed to pose a threat to health.     

Utilization of Industrial Waste for the Production of Bio-Preservative from <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> Me1 and Its Application in Milk and Milk-Based Food Products

The bio-preservative efficacy of a partially purified antibacterial peptide (ppABP) produced by Bacillus licheniformis Me1 in an economical medium developed using agro-industry waste was evaluated by direct application in milk and milk-based food products. The addition of ppABP in milk samples stored at 4 ± 2 °C and 28 ± 2 °C resulted in the growth inhibition of pathogens Listeria monocytogenes Scott A, Micrococcus luteus ATCC 9341, and Staphylococcus aureus FRI 722. The shelf life of milk samples with added ppABP increased to 4 days at 28 ± 2 °C, whereas curdling and off-odor were noticed in samples without ppABP. Furthermore, the milk samples with ppABP were sensorily acceptable. Antilisterial effect was also observed in cheese and paneer samples treated with ppABP. These results clearly indicate that the ppABP of B. licheniformis Me1 can be utilized as a bio-preservative to control the growth of spoilage and pathogenic bacteria, thereby reducing the risk of food-borne diseases.     

Evaluation of Calcium and Magnesium in Scalp Hair Samples of Population Consuming Different Drinking Water: Risk of Kidney Stone

The objective of this study was to examine the relationship between calcium (Ca) and magnesium (Mg) in underground water (UGW), bottled mineral water (BMW), and domestic treated water (DTW) with related to risk of kidney stones. The water samples were collected from different areas of Sindh, Pakistan. The scalp hair samples of both genders, age ranged 30–60 years, consuming different types of water, have or have not kidney disorders, were selected. The Ca and Mg concentrations were determined in scalp hair of study subjects and water by flame atomic absorption spectroscopy. The Ca and Mg contents in different types of drinking water, UGW, DTW, and BMW, were found in the range of 79.1–466, 23.7–140, and 45–270 mg/L and 4.43–125, 5.23–39.6, and 7.16–51.3 mg/L, respectively. It was observed that Ca concentration in the scalp hair samples of kidney stone patients consuming different types of drinking water was found to be higher (2,895–4721 μg/g) while Mg level (84.3–101 μg/g) was lower as compare to referents subjects (2,490–2,730 μg/g for Ca, 107–128 μg/g for Mg) in both genders. The positive correlation was found between Ca and Mg levels in water with related to kidney stone formations in population, especially who consumed underground water. A relative risk and odd ratio were calculated; the relative risk had a strong positive association with incidence of kidney stone which depends on types of drinking water.     

Development of <Emphasis Type="Italic">Cleruchoides noackae</Emphasis>, an egg-parasitoid of <Emphasis Type="Italic">Thaumastocoris peregrinus</Emphasis>, in eggs laid on different substrates,with different ages and post-cold storage

Cleruchoides noackae Lin and Huber (Hymenoptera: Mymaridae) is an egg parasitoid of Thaumastocoris peregrinus Carpintero and Dellapé (Hemiptera: Thaumastocoridae). The parasitism and development of C. noackae was studied in T. peregrinus eggs of different ages, laid on eucalyptus leaves or paper towel and stored at 5 °C. The emergence, sex ratio and development of C. noackae and hatched nymphs of T. peregrinus were evaluated. This parasitoid had an emergence rate higher than 60% from zero to one, one to two, and two to three-day old eggs and lower than 10% for those 3–4 and 4–5 days old. The female proportion was 78% and the egg-adult period for C. noackae was 19.5 days. The use of T. peregrinus eggs up to three days old, laid on paper towel and stored at 5 °C for 14 days did not affect the biological parameters of C. nockae and should be used for mass rearing of this parasitoid.     

Prognostic factors in infective endocarditis in general hospitals in the Netherlands

Introduction

Despite advances in treatment, infective endocarditis (IE) still ranks amongst the most lethal infectious diseases. We sought to determine prognostic factors in general hospitals in the Netherlands as research in this setting is scarce.

Results

Between 2004 and 2011, we identified 216 cases of IE, 30.1?% of which were prosthetic valve IE. This leads to an annual incidence of IE of 5.7 new cases per 100,000 persons per year. Women were less likely to undergo surgical intervention (OR = 1.96, 95?% CI 1.06–3.61, p = 0.031). Also, ageing was an independent prognostic factor for not receiving surgery in a multivariate analysis (annual OR = 1.04, 95?% CI 1.02–1.06, p < 0.001). Female sex was a prognostic factor for mortality (OR = 2.35, 95?% CI 1.29–4.28, p = 0.005). Age was also an independent prognostic factor for mortality (OR = 1.05, 95% CI 1.03–1.08, p < 0.001). Conservative treatment was a prognostic factor for mortality (OR = 3.39, 95?% CI 1.80–6.38, p < 0.001) whereas surgical intervention was an independent prognostic factor for adverse events (OR = 3.03, 95% CI 1.64–5.55, p < 0.001). Staphylococcus aureus was an independent prognostic factor for adverse events (OR = 2.05, 95?% CI 1.10–3.84, p = 0.024) but not for mortality.

Conclusion

This study shows that endocarditis in general hospitals has a high rate of morbidity and mortality. Even when treated, it ranks as one of the most lethal infectious diseases in the Netherlands, especially in women and the elderly.

     

The Concentration of Vanadium in Pathologically Altered Human Kidneys

This study evaluated serum and hair mineral and trace element levels as well as levels of other nutritional factors affecting growth and appetite in young children with non-organic failure to thrive (NOFTT) based on the presence or types of feeding difficulty (FD). Between August 2012 and July 2015, 136 children less than 6 years of age with NOFTT were included. FD was diagnosed based on Wolfson criteria and divided into subtypes according to Chatoor’s classification. Clinical data were reviewed, and serum and hair mineral levels were measured. Of all assessed serum and hair minerals, only hair sulfur contents differed significantly between subjects with and without infantile anorexia (39,392 ± 2211 vs. 40,332 ± 2551 μg/g, P = 0.034). There were no differences in serum and hair mineral levels between children with and without sensory food aversion. Hair copper contents were significantly lower in children with FD of reciprocity (12.3 ± 6.0 vs. 22.4 ± 25.1 μg/g, P = 0.049). While hair zinc contents were also lower, the difference was not statistically significant (49.2 ± 26.8 vs. 70.6 ± 41.0 μg/g, P = 0.055). Only hair manganese contents were significantly lower in children with posttraumatic FD (0.12 ± 0.04 vs. 0.26 ± 0.73 μg/g, P = 0.037). In conclusion, there were no differences in most serum and hair mineral levels in children with NOFTT, except for relatively lower hair levels of sulfur, copper and possibly zinc, and manganese in infantile anorexia, reciprocity, and posttraumatic types of FD, respectively.     

Brucellosis Risk in Urban and Agro-pastoral Areas in Tanzania

Epidemiology of human and animal brucellosis may depend on ecological conditions. A cross-sectional study was conducted to compare prevalence and risk factors of bovine brucellosis, and risky behaviours for the human infection between urban and agro-pastoral areas in Morogoro region, Tanzania. Cattle blood sampling and interviews using a structured questionnaire were conducted with farmers. Rose-Bengal test was conducted for the cattle sera, and positive samples were confirmed with competitive ELISA. Farm-level sero-prevalences were 0.9% (1/106, 95% CI 0.0–5.9%) and 52.9% (9/17, 95% CI 28.5–76.1%) in urban and agro-pastoral areas, respectively. The animal-level-adjusted prevalences were 0.2% (1/667, 95% CI 0.0–1.1%) and 7.0% (28/673, 95% CI 5.7–8.4%) in those areas. The final farm-level model including both areas found two risk factors: history of abortion in the herd (P < 0.01) and cattle grazing (P = 0.07). The animal-level risk factors in agro-pastoral areas were age (P = 0.04) and history of abortion (P = 0.03). No agro-pastoral farmer knew about Brucella vaccine. Agro-pastoralists generally had poorer knowledge on brucellosis and practiced significantly more risky behaviours for human brucellosis such as drinking raw milk (17.6%, P < 0.01) and blood (35.3%, P < 0.01), and helping cattle birth (100%, P = 0.04) than urban farmers (0, 0 and 79.2%, respectively). Intervention programs through education including both human and animal health particularly targeting agro-pastoralists would be needed.     

Molecular characterisation of four echinostomes (Digenea: Echinostomatidae) from birds in New Zealand,with descriptions of <Emphasis Type="Italic">Echinostoma novaezealandense</Emphasis> n. sp. and <Emphasis Type="Italic">Echinoparyphium poulini</Emphasis> n. sp.

Morphological and molecular characterisation of echinostome specimens (Digenea: Echinostomatidae) recovered in one Anas platyrhynchos L. and one Cygnus atratus (Latham) (Anseriformes: Anatidae) from New Zealand revealed the presence of two known species, Echinostoma miyagawai Ishii, 1932 and Echinoparyphium ellisi (Johnston & Simpson, 1944) and two species new to science. Comparative morphological and phylogenetic analyses supported the distinct species status of Echinostoma novaezealandense n. sp. ex Branta canadensis (L.), A. platyrhynchos and C. atratus, and Echinoparyphium poulini n. sp. ex C. atratus. Echinostoma novaezealandense n. sp., a species of the “revolutum” species complex characterised by the possession of a head collar armed with 37 spines, keyed down to E. revolutum but was distinguished from the latter in having a much narrower body with almost parallel margins, longer oesophagus, wider cirrus-sac, larger seminal vesicle, much smaller ventral sucker, ovary, Mehlis’ gland and testes, more anteriorly located ovary and testes, and distinctly smaller eggs (81–87 × 42–53 vs 106–136 × 55–70 µm). This new species appears similar to Echinostoma acuticauda Nicoll, 1914 described in Australia but differs in having a longer forebody, more posteriorly located ovary and testes, and much smaller eggs (81–87 × 42–53 vs 112–126 × 63–75 µm). Echinoparyphium poulini n. sp. is differentiated from the four species of Echinoparyphium possessing 37 collar spines considered valid as follows: from E. chinensis Ku, Li & Chu, 1964 in having a much smaller body, four (vs five) angle spines and simple seminal vesicle (vs bipartite); from E. schulzi Matevosyan, 1951 in having a less robust body at a comparable body length, much smaller ventral sucker, ovary and testes, and longer but narrower eggs (87–109 × 50–59 vs 70–85 × 60–84 µm); and from the two smaller forms, E. serratum Howell, 1968 and E. aconiatum Dietz, 1909, in a number of additional metrical features correlated with body size and especially in the possession of much larger collar spines. Partial fragments of the mitochondrial nad1 and 28S rRNA genes were amplified for representative isolates of the four species and analysed together with sequences for Echinostoma spp. and Echinoparyphium spp. available on GenBank. Phylogenetic analyses based on the mitochondrial nad1 gene revealed congruence between the molecular data and species identification/delineation based on morphology; this was corroborated by the 28S rDNA sequence data.