Single Gall Research Articles

Exploring the diversity of galls on Artemisia indica induced by Rhopalomyia species through morphological and transcriptome analyses.

Plant galls generated by insects have highly organized structures, providing nutrients and shelter to the insects living within them. Most research on the physiological and molecular mechanisms of gall development has focused on single galls. To understand the diversity of gall development, we examined five galls with different morphologies generated by distinct species of Rhopalomyia (gall midge; Diptera: Cecidomyiidae) on a single host plant of Artemisia indica var. maximowiczii (Asteraceae). Vasculature developed de novo within the galls, indicating active transport of nutrients between galls and the host plant. Each gall had a different pattern of vasculature and lignification, probably due to differences in the site of gall generation and the gall midge species. Transcriptome analysis indicated that photosynthetic and cell wall-related genes were down-regulated in leaf and stem galls, respectively, compared with control leaf and stem tissues, whereas genes involved in floral organ development were up-regulated in all types of galls, indicating that transformation from source to sink organs occurs during gall development. Our results help to understand the diversity of galls on a single herbaceous host plant.

Mycobiomes of two distinct clades of ambrosia gall midges (Diptera: Cecidomyiidae) are species-specific in larvae but similar in nutritive mycelia.

Ambrosia gall midges are endophagous insect herbivores whose larvae live enclosed within a single gall for their entire development period. They may exhibit phytomycetophagy, a remarkable feeding mode that involves the consumption of plant biomass and mycelia of their cultivated gall symbionts. Thus, AGMs are ideal model organisms for studying the role of microorganisms in the evolution of host specificity in insects. However, compared to other fungus-farming insects, insect-fungus mutualism in AGMs has been neglected. Our study is the first to use DNA metabarcoding to characterize the complete mycobiome of the entire system of the gall-forming insects as we profiled gall surfaces, nutritive mycelia, and larvae. Interestingly, larval mycobiomes were significantly different from their nutritive mycelia, although Botryosphaeria dothidea dominated the nutritive mycelia, regardless of the evolutionary separation of the tribes studied. Therefore, we confirmed a long-time hypothesized paradigm for the important evolutionary association of this fungus with AGMs.

Incidence, Risk Factors and Outcome of Gallbladder Perforation during Laparoscopic Cholecystectomy

Background and Aim: Gallbladder perforation has been recorded in patients undergoing laparoscopic cholecystectomy and associated with the development of gallstones and bile leakage. The present study investigated the incidence, risk factors, and outcome of gallbladder perforation in patients undergoing elective laparoscopic cholecystectomy. Patients and Methods: This prospective study was conducted on 156 patients who underwent laparoscopic cholecystectomy in the department of General Surgery, Federal Government Polyclinic Hospital, Islamabad from January 2021 to July 2022. Gallbladder stone patients with chronic symptoms such as vomiting, recurrent episodes of epigastric, nausea, and hypochondrial pain on right side were included. Acute cholecystitis patients with single or multiple gallstone disease were also enrolled. All the patients were prepared for laparoscopic cholecystectomy and investigations such as blood sugar, liver function test, CBC, blood urea and creatinine, ultrasound of gallbladder, and ECG were done. SPSS version 26 was used for data analysis. Results: Of the total 156 patients, there were 126 (80.8%) females and 30 (19.2%) male. Age-wise distribution of patients were as follows: 66 (42.3%) in 20-35 years, 52 (33.3%) in 36-50 years, and 38 (24.4%) in 51-65 years. The incidence of single gall stone (SGS), multi gallstones (MGS), and acute cholecystitis was 48 (30.8%), 88 (56.4%), and 20 (12.8%) respectively. The overall mean age was 36 years. The incidence of gallbladder perforation was 26.9% (n=42) among all patients. Patients distribution based on gallbladder perforation were as follows: 17 (10.9%) in MGS, 14 (9.0%) in SGS, and 11 (7.1%) in acute cholecystitis cases. Conclusion: The present study found that the prevalence of gallbladder perforation was 26.9% among SGS, MGS, and acute cholecystitis. Majority of GB perforation occurred during gallbladder bed dissection. Acute cholecystitis patients are more prone to GB perforation. GB perforation can be reduced by utilization of harmonic scalpel in laparoscopic cholecystitis. Keywords: Elective laparoscopic cholecystectomy, Gallbladder perforation, Outcomes

Bifurcation of the cystic duct – A rare reverse “Y” anomaly encountered during laparoscopic cholecystectomy.

Cystic duct anatomy has been quite variable in its presentation, but duplication and bifurcation of the cystic duct draining a single gall bladder is an extremely rare scenario. We present a case of reverse “Y” anomaly of the cystic duct in a 47-year-old male who underwent laparoscopic cholecystectomy (LC). Only a single case of a reverse “Y” cystic duct anomaly was found to be reported in the English literature which was an incidental finding intra-operatively.(1) Anatomical variations being rare are often missed during pre-operative imaging. Careful dissection while delineating the anatomy and proceeding with caution during every laparoscopic cholecystectomy is ideal to avoid ductal injury.

Comparison of Body Mass Index and Cholelithiasis Diagnosed on Ultrasound in the Population of Pindi Bhattian

Obesity is one of the emerging conditions in our population. Gallstones are composed of cholesterol. It is postulated that gallstones are associated with the deposition of cholesterol. Ultrasound is the first line imaging modality for the diagnosis of cholelithiasis. If gallstones are correlated with BMI, it will open up a new vista for further research. In Yaqoob Ultrasound Clinic, PindiBhattian, Pakistan, a cross sectional analytical study was conducted. All included patients were inquired regarding variables like Age, Height, Weight, Presences of stones, Wall thickness of gall bladder, and location of gall stones on grey scale ultrasound findings. Patients were asked to lie down and expose their abdomen; grey scale abdominal ultrasound was performed of these patients. All variables mentioned above for each patient was recorded and maintain in their individual case record form (CFR). Data was collected during allocated period. Statically package SPSS version 2.4 was used to check the collected data and to organize and compile results. According to results out of total 160 patients, 105 were females and 55 were male. Out of 160 patients 150 (93.8%) were present with gall stones and 10 (6.3%) were having no gall stone. In total 61 patients (38.1%) were having single gall stone whereas 89 (55.6%) were diagnosed with multiple gall stones. According to the results, minimum age was 18 years and maximum was 80 years. Our study concluded that obesity is one of the major risk factors for gall stones. Obesity is usually more common in females, so they are more likely to have gall stones and overweight patients were more prone to GB stones.

Gall Bladder Calculus in reproductive age group females attending the ultrasound clinic at a tertiary care hospital: A comparative Cross-sectional study

Background: Pregnant females are at risk for cholelithiasis' progress, and the obstruction in the bile duct produced by gallstones is responsible for jaundice and abdominal pain. Objective: The objectives of the present study were to see the burden of gall stones, relation with parity, and trimester among reproductive age group females. Methodology: A cross-sectional study was conducted on 467 females of age between 15 to 45 years. The gall bladder was thoroughly examined by an ultrasound probe of frequency 2-5 MHz in all planes. Results: In most females who had gallstones, single (68.8 percent) gall stone was common as compared to multiple (31.2 percent). More nonpregnant women had gall bladder stones than pregnant women (p-value <0.005). Among the current nonpregnant group, the percentage of presence of gallbladder stones was greater in the female having parity of two or more (27.4 percent) than the females having parity of less than two (8.2 percent) with p-value <0.05. Conclusion: Since Gallstones can further complicate the situation in the form of gallbladder carcinoma, and females with higher parity have more chances of developing gallstones. Therefore, it becomes necessary to know the trimester of gallstone formation and its role in the progression of symptoms.

The Prevalence and Association between Subclinical Hypothyroidism and Gall Stone Disease: Study in Shaheed Suhrawardy Medical College Hospital

Background: Gall stones are the most common biliary pathology. Subclinical hypothyroidism is not an uncommon problem in the population with thyroid disease, several explanations for a possible relation between hypothyroidism and lipid metabolism with gall stone formation proved that prevalence of gall stones is increased in patients with hypothyroidism.
 Objective: To find the prevalence and correlation between the subclinical hypothyroidism and gall stone disease.
 Methods: This is a cross-sectional study was done in the Unit-II of department of Surgery of Shaheed Suhrawardy Medical Hospital between July 2018 to June 2019, where 80 patients presented with gall stones. All the patients were assessed and prepared for cholecystectomy by detailed history, clinical examination, thyroid function test, abdominal ultrasound and others investigation needed for general anaesthesia.
 Results: Among 80 patients, 78.57% were in 40–59 year age group, 56 (70%) of them were female and 24 (30%) were male. Of the total number of patients, 14 of them (17.5%) had subclinical hypothyroidism and 66 (82.5%) were euthyroid. Most of patients in the subclinical hypothyroid group were showing female gender predominance with 78.57%. While the prevalence among males were found 21.43%. Dyslipidemia was present among 81.25% patients (p=0.03) having subclinical hypothyroidism and gall stones. Besides single stone was common (57.14%) among this hypothyroid group and all stones of this group were >1 cm.
 Conclusion: There is a gender speciûc relationship between subclinical hypothyroidism and gall stone disease. As this study sharing statistically significant in the prevalence of the subclinical hypothyroidism among females in age groupe³40 years, dyslipidemia and single gall stone. This subset of patients should be assessed for thyroid dysfunction.
 J Shaheed Suhrawardy Med Coll, December 2019, Vol.11(2); 130-133

Galhas entomógenas em três fragmentos de Mata Atlântica no extremo Sul Catarinense, Brasil

Fifteen insect gall morphotypes were found on 11 plant species (8 families) in three fragments of the Dense Ombrophylous Lowland Forest, in the municipalities of Maracajá, Içara and Araranguá, Santa Catarina. Most of the galls occurred in leaves, followed by stems galls and a single gall morphotype in Leandra sp. (Melatomastaceae), occurred in both leaves and stems. Glabrous and isolated galls were more frequent than hairy and gregarious galls. Lauraceae was the family of plant with greater wealth of galls. It was pointed out the first record of the plant Microstachys serrulata (Euphorbiaceae) for the state of Santa Catarina. Galls found in Ilex theezans (Aquifoliaceae), Microstachys serrulata (Euphorbiaceae), Dahlstedtia muehlbergiana (Fabaceae), Nectandra oppositifolia (Lauraceae), Leandra sp. (Melastomataceae), Miconia sellowiana (Melastomataceae), Myrcia multiflora (Myrtaceae) and Urera nitida (Urticaceae) were recorded for the first time in the State of Santa Catarina.

Shift of Feeding Mode in an Epizoic Stalked Barnacle Inducing Gall Formation of Host Sea Urchin.

SummaryAmong diverse stalked barnacles, Rugilepas pearsei (Thoracica: Cirripedia: Arthropoda) is a rare unique species that is associated with echinoids and has highly atrophied cirri. We rediscovered the barnacle for the first time from description and verified that the barnacles live obligately in half-open galls formed on the test of the sea urchin Echinothrix diadema (Diadematidae: Echinodermata). A molecular phylogenetic analysis demonstrated that the obligate association with echinoids derived from epizoic life on crustaceans. A stable isotope analysis suggests that the barnacle feeds on particulate organic matter (POM) without parasitizing the host echinoids. These findings suggest that the host shift caused losses of plates and feather-like cirri, changes in the attachment device from cementation to anchoring, and a shift in feeding mode from filter feeding to POM collection. The barnacle's epizoic, superficially sub-endozoic, communal life in stout but narrow galls causes repetitive reproduction at the cost of reduced growth.

Most Plasmodiophora brassicae Populations in Single Canola Root Galls from Alberta Fields are Mixtures of Multiple Strains.

Clubroot, caused by Plasmodiophora brassicae, is an important disease on canola in Alberta, Canada. The pathogen is grouped into pathotypes according to their virulence reaction on differential hosts. Multiple pathotypes or strains are known exist in one field, one plant, or even one gall. This study was conducted with the objective of testing the prevalence of the coexistence of multiple strains in a single gall. In all, 79 canola clubroot galls were collected from 22 fields across Northern Alberta in 2018. Genomic DNA extracted from these single galls was analyzed using RNase H-dependent PCR (rhPCR). The rhPCR primers were designed to amplify a partial sequence of a dimorphic gene, with one primer pair specific to one sequence and the other primer pair specific to the alternative sequence. The amplification of both sequences from DNA obtained from a single gall would indicate that it contains two different P. brassicae strains. The rhPCR analyses indicated that the P. brassicae populations in 50 of the 79 galls consisted of more than one strain. This result emphasizes the need for cautious interpretation of results when a single-gall population is subject to pathotyping or being used as inoculum in plant pathology research. It also confirms that the maintenance of pathotype diversity within single root galls is a common occurrence which has implications for the durability, and stewardship, of single-gene host resistance.

Anatomical forms of Domestic Cat (Felis catus domesticus ) gall bladder in Egypt with its relation to their biliary system

In order to help veterinarians in application of surgical procedures in cat and detection of gall bladder disease so that, we gathered twenty-one domestic cats and concluded that three main forms were collected; single gall bladder, duplex fundus and bilobed gall bladder. Both the gall bladder forms and the hepatic ducts distribution were not related. The bile duct was constructed from the left and right hepatic ducts in addition to the cystic one. In some cases, we explored that a short common hepatic duct. Sonographically, the gall bladder ap- peared differ in conformance, encir- cled by echogenic wall and adjacent to the right medial lobe laterally and the quadrate lobe medially.

Pattern of gall stone (GS) size & number in 1263 otherwise normal Sudanese subjects & their impact in the development of gall bladder disease & complications at the National Centre of Gastroentrology & Hepatology in Sudan during the last 10 years

1263 cases of GS, examined by ultrasound to decide size & number of stones. Size of stones was categorized into sand, tiny(1-4 mm), small(4-1.0 cm), medium(11-20 mm), big(21-30 mm), large(>30 mm), mixed stones(variable sizes). Number of stones categorized into; single, 2 stones, 3 stones, few( 20) & packed(no GB free space). Females scored (74.4%) GS cases. Prevalence of GS significantly increased above the age of 30years. Single stone was the predominant stone (21.2%) followed by multiple (19.6%), least was packed stones(3.1%), number didn't increase significantly with age while size significantly increased with age. Commonest stone size in both sexes was small stone in (32.1%) of cases. Predominance of fine, big & variable GS in females in 81.6%, 85% & 83.7% in total cases respectively. GS with normal gall bladder constituted (55.3%), Acute cholycystitis, had the highest prevalence of (28%) among gall bladder disease, chronic cholycystitis (16.7%). Gall bladder disease had no correlation with stone size & number. Risk of Gall bladder obstruction was significantly increased with single gall stone. Total complications of pericholycystic fluid, empyema, perforation, gangrene, emphysema, pancreatitis, Mirrizi, fistula & mucocoele occured in (9.8%) of cases. The predominant complication was pancreatitis (36.3%) of total complications one third of which had additional pseudopancreatic cyst. Pancreatitis was exclusively related to stone size Conclusion: No correlation of GS size & number to sex, single gall stone was the most prevalent stone number & carried a higher risk of gall bladder obstruction & higher incidence of acute cholycystitis. Acute cholycystitis was the commonest gall bladder disease. Pancreatitis was the most prevalent complication related to Stones

Diversity of galls induced by wasps (Hymenoptera: Cynipidae, Cynipini) associated with oaks (Fagaceae: Quercus ) in Mexico

Background. Gall-inducing insects make up a guild of highly specialized endophagous herbivores. The cynipids (Hymenoptera: Cynipidae: Cynipini) are highly diversified gall-inducing wasps that are largely associated with oaks (Fagaceae: Quercus ). Mexico is one of the centers of diversification for the Quercus genus with 161 described species, of which 109 are endemic.
 Questions / hypothesis. The present study aims to identify the gall richness, gall morphological variation and degree of specificity to oaks in Mexico.
 Methods. An intensive collection was conducted from March to September each year from 2008 to 2012 for a total of 80 oak species in 120 localities in Mexico.
 Results. A total of 224 morphologically distinct galls associated with 73 of the 80 oak species were found. The largest number of morphotypes was found in leaves (125), followed by branches (37), buds (31), petioles (20), catkins (5), acorns (3) and roots (3). The degree of specificity between the gall-inducing wasps and their hosts was highly variable; between one to 20 distinct gall morphotypes were found in each species. Only 23 oak species had a single gall morphotype associated.
 Conclusions. This study demonstrates the important interaction between oaks and gall-inducing wasps, which is a very complex co-evolutionary process. It also shows the relevance of basic taxonomic studies of little-known groups such as gall-inducing wasps, especially in a highly biodiverse country such as Mexico.

Patterns of gall infestation in Heteropterys byrsonimifolia A. Juss. in a forest-savannah ecotone

ABSTRACT Galls are the result of a specific interaction between an inducer and a host plant. The species Heteropterys byrsonimifolia A. Juss. occurs in abundance in semideciduous seasonal forest ecotones and adjacent open formations. In the ecological reserve Quedas do Rio Bonito, located in the state of Minas Gerais, Brazil, this species is affected by a single gall morphotype. The present study aimed to evaluate whether the structural complexity of the host (test of the structural complexity hypothesis) and the distance between hosts (test of the resource concentration hypothesis) affect gall density in H. byrsonimifolia and to characterize the spatial distribution of the infestation. The results corroborate the two hypotheses tested, suggesting a metapopulation pattern of gall infestation in H. byrsonimifolia. Gallers were more successful in abrupt forest-savannah transition environments, which may be associated with greater stress-induced host vulnerability that plants usually experience in ecotones.

Tuberculosis of Gallbladder - A Case Report

Gall bladder is considered a rare site for occurrence of tuberculosis and is usually found in association with gallstones or cystic duct obstruction. The diagnosis is very difficult prior to surgery unless there is strong suspicion of tuberculosis, with the confirmation being possible only after histopathological examination of gall bladder. Here we present a case of gall bladder tuberculosis in a 40-year-old female, who came with symptoms of chronic cholecystitis due to gallstones. Ultrasound of abdomen revealed a single gall stone located in the fundal region. The patient underwent laparoscopic cholecystectomy and was treated with anti-tubercular drugs. .

Co-occurrence of pathogenic and non-pathogenic Fusarium decemcellulare and Lasiodiplodia theobromae isolates in cushion galls disease of cacao (Theobroma cacao L.)

AbstractFlowery cushion gall of cacao is a disease complex with six types. Fusarium decemcellulare have been isolated from both flowery and green point galls and recognized as the etiological agent of the disease. In the present work we: i) identified by ITS-rDNA sequencing and/or taxonomy the cultivable fungal species or Operative Taxonomic Units (OTUs) associated with the five symptoms of cushion galls in cacao from Venezuela, and ii) determined the gall inducing capacity on cacao peeled seeds after 45 days of inoculation with suspensions of mycelia/ spores from distinct isolate types. The whole isolate collection rendered an abundance of 113 isolates with a richness of 39 OTUs (27) and eight identified at the species or genera levels, respectively, and in unidentified fungi. The dominant recovered species (≈36%) were F. decemcellulare and Lasiodiplodia theobromae. Some isolates of F. decemcellulare, L. theobromae, F. equiseti, Fusarium spp., F. solani, F. incarnatum, Rhizocthonia solani and Penicillium sp. were pathogenic. Some other isolates of the first six mentioned taxa behave as non-pathogenic. Furthermore, pathogenic and non-pathogenic isolates can also co-occur within a single plant and gall type. Moreover, 2-5 species within a single gall symptom in a single tree were identified (not necessarily at the same point in the tree), indicating a broad diversity of co-occurring taxa.

Eurytoma serratulae and E. robusta (Hymenoptera, Eurytomidae): complementary host exploitation strategies of coexisting parasitoids and their impact on the host Urophora cardui

Our study investigates the host exploitation strategies of Eurytoma serratulae and E. robusta (Chalcidoidea, Eurytomidae), two parasitoid species that co-occur in gall populations of the tephritid Urophora cardui on Cirsium spp. The endoparasitoid E. serratulae detects the host larvae before an externally visible gall is formed. It profits from large galls, as its parasitization rate increases with increasing numbers of chambers per gall. Oviposition by the ectoparasitoid E. robusta does not occur until a distinct gall with chambers has been formed. Its parasitization rate reaches highest values in medium-sized galls. Eurytoma robusta is the dominant parasitoid in host populations with low and moderate gall densities, whereas E. serratulae is the superior exploiter of host populations with high gall densities. Within single galls E. robusta is an important hyperparasitoid of E. serratulae, but E. serratulae has no adverse influence on E. robusta. Parasitism by E. serratulae induces host larvae to promote gall growth, an effect that is profitable to both the parasitoid and the remaining host larvae in the gall. Parasitism by E. robusta often leads to smaller galls, as cases of unsuccessful parasitization result in empty gall cells.

Insect galls of the xeric vegetation of Ilha do Cabo Frio (Arraial do Cabo, RJ, Brazil)

The xeric vegetation of Ilha do Cabo Frio (Arraial do Cabo, RJ) was investigated for insect galls, in May, June and August of 2012, totalizing 24 hours of field work. Forty-five morphotypes of insect galls were found on 29 species of host plant (25 genera and 18 families). Asteraceae and Myrtaceae were the most galled plant families. Leaves were the most galled plant organ, with ca. 62% of the total, followed by stems (ca. 22%). Globoid and fusiform galls were the most frequent with about 29% and 24%, respectively. The majority was glabrous (ca. 91%), and one-chambered (ca. 84%). Gallers of 30 morphotypes were determined. They belong to Diptera (Cecidomyiidae), and Lepidoptera. The former was the most frequent. Larvae of Muscomorpha (Diptera) (probably inquilines) were found in a single gall morphotype; parasitoids (Hymenoptera) in seven (ca. 15% of the total), and predators (pseudoscorpion) in a single one. As there is no previous data of insect galls of Ilha do Cabo Frio, all records presented here are new. The geographic distribution of nine species of gall midges (Diptera, Cecidomyiidae) is widened. For the first time, eleven plant species are recorded as host of galling insects, and 24 gall morphotypes are described.

SONOGRAPHY TO PREDICT CONVERSION IN LAPAROSCOPIC CHOLECYSTECTOMY

Laparoscopic cholecystectomy has become the gold standard in the treatment of symptomatic cholelithiasis. Several preoperative variables have been identified as risk factors that are helpful in predicting the probability of conversion to laparotomy for safe and successful removal of the gallbladder. This study aimed to look for the various sonographic findings and to make a predictive index for patients who are candidates for laparoscopic cholecystectomy, operative difficulties and conversion to open cholecystectomy. This is a prospective study conducted in the Department of Surgery of Al-Mawani General Hospital, Al-Mousawi Private Hospital in Basrah, Iraq, between May 2005 and October 2008. Abdominal Sonography performed in 105 consecutive patients before laparoscopic cholecystectomy (the sonographic signs are: gallbladder wall thickness, pericholecystic fluid, sonographic Murphy's sign, shrunken gallbladder, number and size of gallstones). Patients excluded are those with history of jaundice, abnormal liver function test, upper abdominal surgery, co-morbid illnesses, extreme obesity, dilated intrahepatic or extrahepatic biliary ducts or those with CBD stones. The surgeon re-evaluates the results of ultrasound with the results obtained during surgery. One hundred and five patients included in the study, 103 patients with gallstones, the other 2 patients having polyps. Ultrasound was accurate 100% in detecting gallstones and polyps, 99 patients (94.3%) have their cholecystectomies via the laparoscope, 75 patient (75.8%) from them show easy procedure, while 24 patients (24.2%) suffered from difficulties. Six patients (5.7%) needed conversion to open cholecystectomy to complete the operation safely. In conclusion, there are many sonographic signs that we can be depend on them to give us an idea about the possibility of conversion to open cholecystectomy, the most specific one is pericholecystic fluid. Secondly gallbladder wall thickness more than 3mm, thirdly, sonographic Murphy's sign, fourthly, shrunken gallbladder, fifthly single gall stone. The other signs are of less specificity like size and multiplicity of gall stones.

Haplotype Inferring via Galled-Tree Networks Is NP-Complete

The problem of determining haplotypes from genotypes has gained considerable prominence in the research community since the beginning of the HapMap project. Here the focus is on determining the sets of SNP values of individual chromosomes (haplotypes), since such information better captures the genetic causes of diseases. One of the main algorithmic tools for haplotyping is based on the assumption that the evolutionary history for the original haplotypes satisfies perfect phylogeny. This tool can be applied only on individual blocks of chromosomes, in which it is assumed that recombinations do not happen. However, exact determination of blocks is usually not possible. It would be desirable to develop a method for haplotyping which can account for recombinations, and thus can be applied on multiblock sections of chromosomes. A natural candidate for such a method is haplotyping via phylogenetic networks (which model recombinations) or their simplified version: galled-tree networks. However, even haplotyping via galled-tree networks appears hard, as the efficient algorithms exist only for very special cases: the galled-tree network has either a single gall or only small galls with two mutations each. Building on our previous results, we show that, in general, haplotyping via galled-tree networks is NP-complete, and it remains NP-complete when galls are allowed to have at most k mutations, for any k ≥ 3.